US11014989B2 - Anti-CLL1 specific single-chain chimeric antigen receptors (scCARs) for cancer immunotherapy - Google Patents

Anti-CLL1 specific single-chain chimeric antigen receptors (scCARs) for cancer immunotherapy Download PDF

Info

Publication number
US11014989B2
US11014989B2 US15/546,633 US201615546633A US11014989B2 US 11014989 B2 US11014989 B2 US 11014989B2 US 201615546633 A US201615546633 A US 201615546633A US 11014989 B2 US11014989 B2 US 11014989B2
Authority
US
United States
Prior art keywords
seq
cll1
cell
domain
epitope1
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Active, expires
Application number
US15/546,633
Other languages
English (en)
Other versions
US20180009895A1 (en
Inventor
Julianne Smith
Julien Valton
Alexandre Juillerat
Philippe Duchateau
Barbra Johnson Sasu
Arvind Rajpal
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Cellectis SA
Original Assignee
Cellectis SA
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Cellectis SA filed Critical Cellectis SA
Publication of US20180009895A1 publication Critical patent/US20180009895A1/en
Assigned to RINAT NEUROSCIENCE CORP. reassignment RINAT NEUROSCIENCE CORP. ASSIGNMENT OF ASSIGNORS INTEREST (SEE DOCUMENT FOR DETAILS). Assignors: RAJPAL, ARVIND, SASU, Barbra Johnson
Assigned to CELLECTIS reassignment CELLECTIS ASSIGNMENT OF ASSIGNORS INTEREST (SEE DOCUMENT FOR DETAILS). Assignors: DUCHATEAU, PHILIPPE, VALTON, JULIEN, JUILLERAT, ALEXANDRE, SMITH, JULIANNE
Assigned to CELLECTIS reassignment CELLECTIS ASSIGNMENT OF ASSIGNORS INTEREST (SEE DOCUMENT FOR DETAILS). Assignors: RINAT NEUROSCIENCE CORP.
Application granted granted Critical
Publication of US11014989B2 publication Critical patent/US11014989B2/en
Active legal-status Critical Current
Adjusted expiration legal-status Critical

Links

Images

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
    • A61K35/14—Blood; Artificial blood
    • A61K35/17—Lymphocytes; B-cells; T-cells; Natural killer cells; Interferon-activated or cytokine-activated lymphocytes
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/0005—Vertebrate antigens
    • A61K39/0011—Cancer antigens
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/0005—Vertebrate antigens
    • A61K39/0011—Cancer antigens
    • A61K39/001176—Heat shock proteins
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
    • A61K39/39533—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum against materials from animals
    • A61K39/39558—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum against materials from animals against tumor tissues, cells, antigens
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K40/00—Cellular immunotherapy
    • A61K40/10—Cellular immunotherapy characterised by the cell type used
    • A61K40/11—T-cells, e.g. tumour infiltrating lymphocytes [TIL] or regulatory T [Treg] cells; Lymphokine-activated killer [LAK] cells
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K40/00—Cellular immunotherapy
    • A61K40/30—Cellular immunotherapy characterised by the recombinant expression of specific molecules in the cells of the immune system
    • A61K40/31—Chimeric antigen receptors [CAR]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K40/00—Cellular immunotherapy
    • A61K40/30—Cellular immunotherapy characterised by the recombinant expression of specific molecules in the cells of the immune system
    • A61K40/32—T-cell receptors [TCR]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K40/00—Cellular immunotherapy
    • A61K40/40—Cellular immunotherapy characterised by antigens that are targeted or presented by cells of the immune system
    • A61K40/41—Vertebrate antigens
    • A61K40/42—Cancer antigens
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K40/00—Cellular immunotherapy
    • A61K40/40—Cellular immunotherapy characterised by antigens that are targeted or presented by cells of the immune system
    • A61K40/41—Vertebrate antigens
    • A61K40/42—Cancer antigens
    • A61K40/4202—Receptors, cell surface antigens or cell surface determinants
    • A61K40/4214—Receptors for cytokines
    • A61K40/4215—Receptors for tumor necrosis factors [TNF], e.g. lymphotoxin receptor [LTR], CD30
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K40/00—Cellular immunotherapy
    • A61K40/40—Cellular immunotherapy characterised by antigens that are targeted or presented by cells of the immune system
    • A61K40/41—Vertebrate antigens
    • A61K40/42—Cancer antigens
    • A61K40/4202—Receptors, cell surface antigens or cell surface determinants
    • A61K40/4214—Receptors for cytokines
    • A61K40/4217—Receptors for interleukins [IL]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A61P35/02—Antineoplastic agents specific for leukemia
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
    • C07K14/70503—Immunoglobulin superfamily
    • C07K14/7051—T-cell receptor (TcR)-CD3 complex
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
    • C07K14/70503—Immunoglobulin superfamily
    • C07K14/70517—CD8
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
    • C07K14/70503—Immunoglobulin superfamily
    • C07K14/70535—Fc-receptors, e.g. CD16, CD32, CD64 (CD2314/705F)
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
    • C07K14/70578—NGF-receptor/TNF-receptor superfamily, e.g. CD27, CD30, CD40, CD95
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
    • C07K14/70596—Molecules with a "CD"-designation not provided for elsewhere
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C07K16/2803—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C07K16/2866—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against receptors for cytokines, lymphokines, interferons
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C07K16/2887—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against CD20
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C07K16/30—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants from tumour cells
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C07K16/30—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants from tumour cells
    • C07K16/3061—Blood cells
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K19/00—Hybrid peptides, i.e. peptides covalently bound to nucleic acids, or non-covalently bound protein-protein complexes
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
    • C12N5/0081—Purging biological preparations of unwanted cells
    • C12N5/0093—Purging against cancer cells
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
    • C12N5/06—Animal cells or tissues; Human cells or tissues
    • C12N5/0602—Vertebrate cells
    • C12N5/0634—Cells from the blood or the immune system
    • C12N5/0636—T lymphocytes
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
    • C12N5/06—Animal cells or tissues; Human cells or tissues
    • C12N5/0602—Vertebrate cells
    • C12N5/0634—Cells from the blood or the immune system
    • C12N5/0636—T lymphocytes
    • C12N5/0638—Cytotoxic T lymphocytes [CTL] or lymphokine activated killer cells [LAK]
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
    • C12N5/10—Cells modified by introduction of foreign genetic material
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
    • G01N15/10—Investigating individual particles
    • G01N15/1031—Investigating individual particles by measuring electrical or magnetic effects
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
    • G01N15/10—Investigating individual particles
    • G01N15/14—Optical investigation techniques, e.g. flow cytometry
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/505—Medicinal preparations containing antigens or antibodies comprising antibodies
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/51—Medicinal preparations containing antigens or antibodies comprising whole cells, viruses or DNA/RNA
    • A61K2039/515—Animal cells
    • A61K2039/5156—Animal cells expressing foreign proteins
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/51—Medicinal preparations containing antigens or antibodies comprising whole cells, viruses or DNA/RNA
    • A61K2039/515—Animal cells
    • A61K2039/5158—Antigen-pulsed cells, e.g. T-cells
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/60—Medicinal preparations containing antigens or antibodies characteristics by the carrier linked to the antigen
    • A61K2039/6031—Proteins
    • A61K2039/6056—Antibodies
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/64—Medicinal preparations containing antigens or antibodies characterised by the architecture of the carrier-antigen complex, e.g. repetition of carrier-antigen units
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2239/00—Indexing codes associated with cellular immunotherapy of group A61K40/00
    • A61K2239/10—Indexing codes associated with cellular immunotherapy of group A61K40/00 characterized by the structure of the chimeric antigen receptor [CAR]
    • A61K2239/21—Transmembrane domain
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2239/00—Indexing codes associated with cellular immunotherapy of group A61K40/00
    • A61K2239/10—Indexing codes associated with cellular immunotherapy of group A61K40/00 characterized by the structure of the chimeric antigen receptor [CAR]
    • A61K2239/22—Intracellular domain
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/20—Immunoglobulins specific features characterized by taxonomic origin
    • C07K2317/24—Immunoglobulins specific features characterized by taxonomic origin containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
    • C07K2317/52—Constant or Fc region; Isotype
    • C07K2317/53—Hinge
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
    • C07K2317/56—Immunoglobulins specific features characterized by immunoglobulin fragments variable (Fv) region, i.e. VH and/or VL
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/60—Immunoglobulins specific features characterized by non-natural combinations of immunoglobulin fragments
    • C07K2317/62—Immunoglobulins specific features characterized by non-natural combinations of immunoglobulin fragments comprising only variable region components
    • C07K2317/622—Single chain antibody (scFv)
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/01—Fusion polypeptide containing a localisation/targetting motif
    • C07K2319/02—Fusion polypeptide containing a localisation/targetting motif containing a signal sequence
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/01—Fusion polypeptide containing a localisation/targetting motif
    • C07K2319/03—Fusion polypeptide containing a localisation/targetting motif containing a transmembrane segment
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/33—Fusion polypeptide fusions for targeting to specific cell types, e.g. tissue specific targeting, targeting of a bacterial subspecies
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2510/00—Genetically modified cells
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
    • G01N15/10—Investigating individual particles
    • G01N15/14—Optical investigation techniques, e.g. flow cytometry
    • G01N15/149—Optical investigation techniques, e.g. flow cytometry specially adapted for sorting particles, e.g. by their size or optical properties
    • G01N2015/008—
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
    • G01N15/01—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials specially adapted for biological cells, e.g. blood cells
    • G01N2015/016—White blood cells
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
    • G01N15/10—Investigating individual particles
    • G01N2015/1006—Investigating individual particles for cytology
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
    • G01N15/10—Investigating individual particles
    • G01N2015/1028—Sorting particles
    • G01N2015/1081—
    • G01N2015/149—

Definitions

  • CLL1 C-type lectin-like molecule-1
  • AML acute myeloid leukemia
  • LSC normal hematopoietic stem cells or other cell types.
  • the present invention relates to methods to target CLL1 positive malignant cells using Chimeric Antigen Receptors (anti CLL1-CAR), which are recombinant chimeric proteins able to redirect immune cell specificity and reactivity toward selected membrane antigen CLL1.
  • anti-CLL1 CAR more particularly comprise an extracellular ligand binding comprising a scFV derived from some specific anti-CLL1 monoclonal antibodies.
  • the engineered immune cells endowed with such CARs confer adoptive immunity against CLL1 positive cell as part as various cell therapies for treating cancer, in particular hematologic cancers, with higher efficiency.
  • scCARs transgenic T cell receptors or chimeric antigen receptors
  • scCARs are synthetic receptors consisting of a targeting moiety that is associated with one or more signaling domains in a single fusion molecule.
  • the binding moiety of a scCAR consists of an antigen-binding domain of a single-chain antibody (scFv), comprising the light and variable fragments of a monoclonal antibody joined by a flexible linker. Binding moieties based on receptor or ligand domains have also been used successfully.
  • the signaling domains for first generation scCARs are derived from the cytoplasmic region of the CD3zeta or the Fc receptor gamma chains.
  • First generation scCARs have been shown to successfully redirect T cell cytotoxicity, however, they failed to provide prolonged expansion and anti-tumor activity in vivo.
  • Signaling domains from co-stimulatory molecules including CD28, OX-40 (CD134), and 4-1BB (CD137) have been added alone (second generation) or in combination (third generation) to enhance survival and increase proliferation of scCAR modified T cells.
  • scCARs have successfully allowed T cells to be redirected against antigens expressed at the surface of tumor cells from various malignancies including lymphomas and solid tumors (Jena, Dotti et al. 2010).
  • AML acute myeloid leukemia
  • induction treatments for acute myeloid leukemia have remained largely unchanged for nearly 50 years and AML remains a disease of poor prognosis.
  • AML is a disease characterized by the rapid proliferation of immature myeloid cells in the bone marrow resulting in dysfunctional hematopoiesis.
  • standard induction chemotherapy can induce complete remissions, many patients eventually relapse and succumb to the disease, calling for the development of novel therapeutics for AML.
  • Recent advances in the immunophenotyping of AML cells have revealed several AML associated cell surface antigens that may act as targets for future therapies.
  • CLL1 C-Type Lectin-Like Molecule-1
  • CLL1 appears to be an interesting tumoral antigen target as it is expressed by leukemic blasts at diagnosis from 85-92% of AML patients analysed It is a 75 kDa member of the group V C-type lectin-like receptor family of molecules. Group V molecules have a lectin-like domain that binds to non-sugar ligands.
  • CLL1 is a 265 amino acid type II transmembrane glycoprotein (Uniprot database: Q5QGZ9 for human protein encoded by gene no 160364 in “Entrez Gene” database) that contains a 200 AA extracellular domain.
  • CLL1 is also referred to in the literature and databases as MICL, CLEC12 and KLRL1.
  • CLL1 antigen is associated with AML stem cells.
  • CLL1 is a cell surface protein that is specifically expressed on most malignant lymphoid stem cells (AML LSC), while not being expressed on normal HSC (Van Rhenen et al, 2007). Meanwhile, CLL1 was revealed to be a diagnostic marker in AML (Larsen et al, 2012).
  • Anti-CLL-1 antibodies enable both AML-specific stem-cell detection and possibly antigen-targeting as distinguishing malignant cells from normal stem cells both at diagnosis and in remission (van Rhenen et al, 2007). However, none of these antibodies have been reported to date as being tested in clinical trials as therapeutic antibodies.
  • Gemtuzumab ozogamicin (Mylotarg®) is a monoclonal antibody with a cell poison attached to it. Previously approved to treat AML in older patients, it was withdrawn from the market after studies found some toxicity associated with the product (press release of Dec. 10, 2010 in PMLIVE “ASH: Pfizer eyes re-launch of Mylotarg”). Other monoclonal therapeutic antibodies have shown adverse effects over the last decade (Klastersky, J. (2006) “Adverse effects of the humanized antibodies used as cancer therapeutics” Current Opinion in Oncology. 18(4):316-320)
  • micellar nanoparticles covalently decorated with CLL1-targeting peptides have been described for targeted drug delivery (daunorubicin); these “targeting nanomicelles” transport the drug load to the interior of cells expressing CLL1 and to LSCs isolated from clinical specimens in vitro. It was showed that CLL1-targeting nanomicelles had the potential to be used for targeted drug delivery to leukemia stem cells. However, no therapeutic effects could be attributed to the CCL-1 targeting peptide per se.
  • the inventors have pursued a new approach to target CLL1 using immune cells endowed with specific chimeric antigen receptors based on anti-CLL1 monoclonal antibodies, which redirect immune cell specificity towards CLL1 positive cell.
  • the engineered immune cells that they obtained using this approach have proven efficacy to eliminate CLL1 positive malignant cells.
  • they have appeared to be particularly useful in the context of the production of allogeneic TCR negative engineered immune cells, allowing a reduction of side effects, such as GvHD.
  • the present invention opens the way to treating patients affected with a condition characterized by an overabundance of CLL1-expressing cells using adoptive immunotherapy. Even more, the present invention provides with engineered allogeneic immune cells that may be used as “off-the-shelf” allogeneic therapeutic products. As a further advantage of the invention, the CAR positive engineered cells can be made compatible (i.e. resistant) with chemotherapy or immunodepleting treatments, thereby enabling synergistic effects between chemotherapy and immunotherapy.
  • the inventors have generated CLL1 specific single-chain scCAR having different design and comprising different scFV derived from anti-CLL1 specific antibodies.
  • the Inventors have developed anti-CLL1 specific single-chain CAR (scCAR) comprising VL and VL chains derived from SC02-357, SC02-378, SCO2-161, M26, M31, G4, M22, M29, M2, M5, G12, 21.26 and 1075.7 antibodies, with different architectures and identified highly specific and very selective scCARs constructions that bind to CLL1 expressing cells and selectively destroy CLL1 expressing cancer cells.
  • scCAR anti-CLL1 specific single-chain CAR
  • T-cells from donors have been transformed with polynucleotides expressing these scCARs using viral transduction.
  • the T-cells were further engineered to create less or non-alloreactive T-cells, more especially by disruption of a component of TCR ( ⁇ -T-Cell receptors) to prevent Graft versus host reaction.
  • T-cells were further engineered to create T cells resistant to anti-cancer drugs, to be used in combination with said classical anti-cancer drugs.
  • the resulting engineered T-cells displayed reactivity in-vitro against CLL1 positive cells to various extend, showing that the scCARs of the present invention contribute to antigen dependent activation, and also proliferation, of the T-cells, making them useful for immunotherapy.
  • the resulting engineered T-cells displayed reactivity in-vivo against CLL1 positive cells and significantly reduce the number of cancer cells in vivo.
  • the engineered T-cells of the invention are designed to display in-vivo reactivity against CLL1 positive cells, can be used in concomitance with anti-cancer drugs, are well tolerated. In a particular embodiment, the engineered T-cells of the invention remain efficient even after several administrations, making them useful for immunotherapy as a first treatment (induction), as a consolidation treatment, as a treatment in combination with classical anticancer chemotherapy.
  • the polypeptides and polynucleotide sequences encoding the CARs of the present invention are detailed in the present specification.
  • the engineered immune cells of the present invention are particularly useful for therapeutic applications such as acute myeloma leukemia (AML) treatments.
  • AML acute myeloma leukemia
  • FIG. 1 Schematic representation of an engineered immune cell according to the invention.
  • the engineered immune cell presented in this figure is a T-cell transduced with a retroviral vector encoding CLL1-scCAR.
  • This T-cell was further engineered to allow a better and safer engraftment into the patient, which is optional within the frame of the present invention.
  • X gene may be for instance a gene expressing a component of TCR (TCRalpha or TCRbeta)
  • Y may be a gene involved into the sensitivity of T-cells to immune-suppressive drugs like CD52 (with respect to Campath) or HPRT (with respect to 6-Thioguanine).
  • FIG. 2 Schematic representation of the different scCAR Architecture (V 1 to V6) of the invention (anti-CLL1 scCAR) with the components presented in the following Table 1.
  • FIG. 3A and FIG. 3B Schematic representation of exemplary CLL1 specific CARs according to the invention involving different mAb-epitope tagging for T cell depletion, especially CD20 mimotope(s), which are designed to mitigate possible side effects associated with CAR positive cells injection.
  • V 1 and v2 represents either VH or VL chain respectively of an antibody binding CLL1, TM: transmembrane domain, L: linker, TM: Transmembrane domain (preferably CD8 ⁇ transmembrane domain), 4-1BB: intracellular co-stimulatory domain, CD3 ITAM: activation domain.
  • C CLL1 specific CAR architectures according to the invention, where two epitopes are inserted in the extracellular ligand binding domain of the CAR, one is inserted between the N-terminal end of the CAR and the VH chain, said epitope being bordered by at least one or two linkers; the second epitope is inserted between the VH and VL chains, said 2 nd epitope being also bordered by 2 at least one or two linkers.
  • the architectures illustrated herein differ by the linkers used bordering the 2 nd epitope.
  • the architectures illustrated herein differ by the linkers used bordering the 1 st epitope.
  • These two architectures differ by the linkers used bordering the 2 nd epitope.
  • epitopes preferably CD20 epitopes
  • one CD34 epitope is included between two CD20 epitopes.
  • Further architectures may be considered where CD34 replaces any other previous CD20 epitopes.
  • VFLFPPKPKDTLMIARTPEVTCV VVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRVVSVLTV LHQDWLNGKEYKCKVSNKALPAP IEKTISKAKGQPREPQVYTLPPS RDELTKNQVSLTCLVKGFYPSDI AVEWESNGQPENNYKTTPPVLDS DGSFFLYSKLTVDKSRWQQGNVF SCSVMHEALHNHYTQKSLSLSPG K CD8 ⁇ SEQ ID IYIWAPLAGTCGVLLLSLVITLY transmembrane NO. 6 C domain 41BB SEQ ID IISFFLALTSTALLFLLFFLTLR transmembrane NO.
  • WIGEIYHSGSPNYNPSLKSRVTISVDKSKNQFSLKLSSVTAADTAVYYCAR QTTAGSFDYWGQGTLVTVSS CDR1 SEQ ID GSISSSNWWS NO. 125 CDR2 SEQ ID WIGEIYHSGSPNY NO. 126 CDR3 SEQ ID RQTTAGSFDY NO. 127 SC02-357 & SC02-378 & SEQ ID DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYA SC02-161 light chain NO.
  • V1-SCO2-161 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID scCAR (SEQ ID NO. 1 NO. 13 NO. 14 NO. 3 NO. 6 NO. 8 NO. 9 NO. 47)
  • V1-M26 scCAR SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID (SEQ ID NO. 53) NO. 1 NO. 15 NO. 16 NO. 3 NO. 6 NO. 8 NO. 9
  • V2-SCO2-161 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID scCAR NO. 1 NO. 13 NO. 14 NO. 3 NO. 7 NO. 8 NO. 9 (SEQ ID NO. 48) V2-M26 scCAR SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID (SEQ ID NO. 54) NO. 1 NO. 15 NO. 16 NO. 3 NO. 7 NO. 8 NO. 9 V2-M31 scCAR SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID (SEQ ID NO. 60) NO. 1 NO. 17 NO. 18 NO. 3 NO. 7 NO. 8 NO.
  • V3-SCO2-161 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID scCAR NO. 1 NO. 13 NO. 14 NO. 4 NO. 6 NO. 8 NO. 9 (SEQ ID NO. 49) V3-M26 scCAR SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID (SEQ ID NO. 55) NO. 1 NO. 15 NO. 16 NO. 4 NO. 6 NO. 8 NO. 9 V3-M31 scCAR SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID (SEQ ID NO. 61) NO. 1 NO. 17 NO. 18 NO. 4 NO. 6 NO. 8 NO.
  • V-4 scCAR scCAR Structure Designation signal V-4 peptide VH VL CD8 ⁇ hinge 41BB-TM 41BB-IC CD3 ⁇ CD V4-SCO2-357 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID scCAR NO. 1 NO. 11 NO. 14 NO. 4 NO. 6 NO. 8 NO. 9 (SEQ ID NO. 38) V4-SCO2-378 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID scCAR (SEQ ID NO. 1 NO. 12 NO. 14 NO. 4 NO. 6 NO. 8 NO. 9 NO.
  • V4-SCO2-161 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID scCAR NO. 1 NO. 13 NO. 14 NO. 4 NO. 6 NO. 8 NO. 9 (SEQ ID NO. 50) V4-M26 scCAR SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID (SEQ ID NO. 56) NO. 1 NO. 16 NO. 14 NO. 4 NO. 6 NO. 8 NO. 9 V4-M31 scCAR SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID (SEQ ID NO. 62) NO. 1 NO. 17 NO. 18 NO. 4 NO. 7 NO. 8 NO.
  • V5-SCO2-161 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID scCAR NO. 1 NO. 13 NO. 14 NO. 5 NO. 6 NO. 8 NO. 9 (SEQ ID NO. 51) V5-M26 scCAR SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID (SEQ ID NO. 57) NO. 1 NO. 15 NO. 16 NO. 5 NO. 6 NO. 8 NO. 9 V5-M31 scCAR SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID (SEQ ID NO. 63) NO. 1 NO. 17 NO. 18 NO. 5 NO. 6 NO. 8 NO.
  • V6-SCO2-161 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID scCAR NO. 1 NO. 13 NO. 14 NO. 5 NO. 7 NO. 8 NO. 9 (SEQ ID NO. 52) V6-M26 scCAR SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID (SEQ ID NO. 58) NO. 1 NO. 15 NO. 16 NO. 5 NO. 7 NO. 8 NO. 9 V6-M31 scCAR SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID (SEQ ID NO. 64) NO. 1 NO. 17 NO. 18 NO. 5 NO. 7 NO. 8 NO.
  • the present invention provides:
  • the present invention relates to CLL1 specific chimeric antigen receptor comprising an extracellular ligand-binding domain specifically directed against one portion of the CLL1 antigen, a transmembrane domain and a signaling transducing domain.
  • chimeric antigen receptor is intended molecules that combine an extracellular binding domain directed against a component present on a target cell, for example an antibody-based specificity for a desired antigen (e.g., tumor antigen) with an immune cell receptor component to generate a chimeric protein that will transduce an activating or inhibitory signal toward cellular immune activity.
  • a desired antigen e.g., tumor antigen
  • the present invention more particularly relates to a CLL1 specific chimeric antigen receptor (anti-CLL1 CAR) comprising at least:
  • the CLL1 specific chimeric antigen receptor according to the invention further comprises a co-stimulatory domain, and more preferably a CD28 or a 4-1BB co-stimulatory domain as described for instance by Jena, B., G. Dotti, et al. (2010). It can also comprise a transmembrane domain which can be a Cd8 ⁇ transmembrane domain, as well as an optional hinge.
  • the signal transducing domain or “cytoplasmic signaling domain” of a CAR is responsible for intracellular signaling following the binding of extracellular ligand binding domain to the target resulting in the activation or inhibition of the immune cell and immune response.
  • the signal transducing domain is responsible for the activation or inactivation of at least one of the normal effector functions of the immune cell in which the CAR is expressed.
  • the effector function of a T cell can be a cytolytic activity or helper activity including the secretion of cytokines.
  • the term “cytoplasmic signaling domain” refers to the portion of a protein which transduces the effector signal function signal and directs the cell to perform a specialized function.
  • the cytoplasmic signaling domain which is preferably from a human protein involved in signal transduction pathway(s), determines whether anti-CLL1 CAR is a positive CAR (PCAR) or a negative CAR (NCAR) depending on the nature of the signaling. Respectively, the CAR is a PCAR when the signaling domain, such as CD3zeta from human TCR receptor, has the effect of stimulating the cellular immune activity of the immune cell when the extracellular ligand binding-domain is bound to CLL1.
  • PCAR positive CAR
  • NCAR negative CAR
  • the anti-CLL1 CAR is a NCAR or inhibitory CAR (iCAR) when the signaling domain has the effect of reducing the cellular immune activity, such as signaling domains of human immunoinhibitory receptors CTLA-4 and PD-1 (Federov et al., Sci Transl Med. 2013 Dec. 11; 5 (215): 215ra172).
  • iCAR inhibitory CAR
  • Preferred examples of signal transducing domain for use in a anti-CLL1 CAR can be the cytoplasmic sequences of the T cell receptor and co-receptors that act in concert to initiate signal transduction following antigen receptor engagement, as well as any derivate or variant of these sequences and any synthetic sequence that has the same functional capability.
  • Signal transduction domain comprises two distinct classes of cytoplasmic signaling sequence, those that initiate antigen-dependent primary activation, and those that act in an antigen-independent manner to provide a secondary or co-stimulatory signal.
  • Primary cytoplasmic signaling sequence can comprise signaling motifs which are known as immunoreceptor tyrosine-based activation motifs of ITAMs.
  • ITAMs are well defined signaling motifs found in the intracytoplasmic tail of a variety of receptors that serve as binding sites for syk/zap70 class tyrosine kinases.
  • the signaling transducing domain of the anti-CLL1 CAR can comprise the CD3zeta signaling domain which has amino acid sequence with at least 70%, preferably at least 80%, more preferably at least 90%, 95% 97% or 99% or 100% sequence identity with amino acid sequence selected from the group consisting of SEQ ID NO: 9.
  • a co-stimulatory molecule is a cell surface molecule other than an antigen receptor or their ligands that is required for an efficient immune response.
  • “Co-stimulatory ligand” refers to a molecule on an antigen presenting cell that specifically binds a cognate co-stimulatory molecule on a T-cell, thereby providing a signal which, in addition to the primary signal provided by, for instance, binding of a TCR/CD3 complex with an MHC molecule loaded with peptide, mediates a T cell response, including, but not limited to, proliferation activation, differentiation and the like.
  • a co-stimulatory ligand can include but is not limited to CD7, B7-1 (CD80), B7-2 (CD86), PD-L1, PD-L2, 4-1BBL, OX40L, inducible co-stimulatory ligand (ICOS-L), intercellular adhesion molecule (ICAM, CD30L, CD40, CD70, CD83, HLA-G, MICA, M1CB, HVEM, lymphotoxin beta receptor, 3/TR6, ILT3, ILT4, an agonist or antibody that binds Toll ligand receptor and a ligand that specifically binds with B7-H3.
  • a co-stimulatory ligand also encompasses, inter alia, an antibody that specifically binds with a co-stimulatory molecule present on a T cell, such as but not limited to, CD27, CD28, 4-1BB, OX40, CD30, CD40, PD-1, ICOS, lymphocyte function-associated antigen-1 (LFA-1), CD2, CD7, LTGHT, NKG2C, B7-H3, a ligand that specifically binds with CD83.
  • a “co-stimulatory molecule” refers to the cognate binding partner on a T-cell that specifically binds with a co-stimulatory ligand, thereby mediating a co-stimulatory response by the cell, such as, but not limited to proliferation.
  • Co-stimulatory molecules include, but are not limited to an MHC class I molecule, BTLA and Toll ligand receptor.
  • costimulatory molecules include CD27, CD28, CD8, 4-1BB (CD137), OX40, CD30, CD40, PD-1, ICOS, lymphocyte function-associated antigen-1 (LFA-1), CD2, CD7, LIGHT, NKG2C, B7-H3 and a ligand that specifically binds with CD83 and the like.
  • the co-stimulatory domain of the anti-CLL1 CAR of the present invention comprises a part of co-stimulatory signal molecule selected from the group consisting of fragment of 4-1BB (GenBankTM: AAA53133.) and CD28 (NP_006130.1).
  • the signal transduction domain of the anti-CLL1 CAR of the present invention comprises amino acid sequence which comprises at least 70%, preferably at least 80%, more preferably at least 90%, 95% 97% or 99% sequence identity with amino acid sequence selected from the group consisting of SEQ ID NO: 8.
  • An anti-CLL1 CAR generally further comprises a transmembrane domain (TM).
  • TM transmembrane domain
  • the distinguishing features of appropriate transmembrane domains comprise the ability to be expressed at the surface of a cell, preferably in the present invention an immune cell, in particular lymphocyte cells or Natural killer (NK) cells, and to interact together for directing cellular response of immune cell against a predefined target cell.
  • the transmembrane domain can be derived either from a natural or from a synthetic source.
  • the transmembrane domain can be derived from any membrane-bound or transmembrane protein.
  • the transmembrane polypeptide can be a subunit of the T-cell receptor such as ⁇ , ⁇ , ⁇ or ⁇ , polypeptide constituting CD3 complex, IL2 receptor p55 ( ⁇ chain), p75 (( ⁇ chain) or ⁇ chain, subunit chain of Fc receptors, in particular Fc ⁇ receptor III or CD proteins.
  • the transmembrane domain can be synthetic and can comprise predominantly hydrophobic residues such as leucine and valine.
  • said transmembrane domain is derived from the human CD8 alpha chain (e.g. NP_001139345.1)
  • the transmembrane domain can further comprise a hinge region between said extracellular ligand-binding domain and said transmembrane domain.
  • hinge region generally means any oligo- or polypeptide that functions to link the transmembrane domain to the extracellular ligand-binding domain.
  • hinge region are used to provide more flexibility and accessibility for the extracellular ligand-binding domain.
  • a hinge region may comprise up to 300 amino acids, preferably 10 to 100 amino acids and most preferably 25 to 50 amino acids.
  • Hinge region may be derived from all or part of naturally occurring molecules, such as from all or part of the extracellular region of CD8, CD4 or CD28, or from all or part of an antibody constant region.
  • the hinge region may be a synthetic sequence that corresponds to a naturally occurring hinge sequence, or may be an entirely synthetic hinge sequence.
  • said hinge domain comprises a part of human CD8 alpha chain, Fc ⁇ RIII ⁇ receptor or IgG1 respectively referred to in this specification as SEQ ID NO. 3, SEQ ID NO. 4 and SEQ ID NO.5, or hinge polypeptides which display preferably at least 80%, more preferably at least 90%, 95% 97% or 99% sequence identity with these polypeptides.
  • the hinge can also be a human Ig (immunoglobulin) hinge, e.g., a PD-1 hinge, an IgG4 hinge.
  • the anti-CLL1 CAR according to the invention comprises a transmembrane domain more particularly selected from Cd8 ⁇ and 4-1BB, showing identity with the polypeptides of SEQ ID NO. 6 or 7.
  • An anti-CLL1 CAR according to the invention generally further comprises a transmembrane domain (TM) more particularly a TM selected from Cd8 ⁇ and 4-1BB, and even more particularly showing identity with the polypeptides of SEQ ID NO. 6 or 7.
  • TM transmembrane domain
  • a anti-CLL1 CAR according to the invention further comprises a TM domain from Cd8 ⁇ with SEQ ID NO. 6 or showing at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity with SEQ ID NO. 6
  • the CLL1 specific anti-CLL1 CAR according to the invention can comprise another extracellular ligand-binding domains, to simultaneously bind different elements in target thereby augmenting immune cell activation and function.
  • the extracellular ligand-binding domains can be placed in tandem on the same transmembrane polypeptide, and optionally can be separated by a linker.
  • said different extracellular ligand-binding domains can be placed on different transmembrane polypeptides composing the anti-CLL1 CAR.
  • the present invention relates to a population of anti-CLL1 CAR s comprising each one different extracellular ligand binding domains.
  • the present invention relates to a method of engineering immune cells comprising providing an immune cell and expressing at the surface of said cell a population of anti-CLL1 CAR each one comprising different extracellular ligand binding domains.
  • the present invention relates to a method of engineering an immune cell comprising providing an immune cell and introducing into said cell polynucleotides encoding polypeptides composing a population of anti-CLL1 CAR each one comprising different extracellular ligand binding domains.
  • population of anti-CLL1 CAR s it is meant at least two, three, four, five, six or more anti-CLL1 CAR s each one comprising different extracellular ligand binding domains.
  • the different extracellular ligand binding domains according to the present invention can preferably simultaneously bind different elements in target thereby augmenting immune cell activation and function.
  • the present invention also relates to an isolated immune cell which comprises a population of anti-CLL1 CAR s each one comprising different extracellular ligand binding domains.
  • CLL1 specific chimeric antigen receptors can have different architectures, as they can be expressed, for instance, under a single-chain chimeric protein (scCAR) or under the form of several polypeptides (multi-chain) including at least one such chimeric protein.
  • scCAR single-chain chimeric protein
  • multi-chain multi-chain CAR architectures are disclosed in WO2014/039523, especially in FIGS. 2 to 4, and from page 14 to 21, which are herein incorporated by reference.
  • anti-CLL1 CAR comprises an extracellular single chain antibody (scFv Fc) fused to the intracellular signaling domain of T-cell antigen receptor complex zeta chain (scFv Fc: ⁇ ), which has the ability, when expressed in T cells, to redirect antigen recognition based on the monoclonal antibody's specificity.
  • scFv Fc extracellular single chain antibody fused to the intracellular signaling domain of T-cell antigen receptor complex zeta chain
  • the present application discloses several anti-CLL1 single chain CAR directed against CLL1 antigen, which comprise as non-limiting example the amino acid sequences: SEQ ID NO: 35 to 112.
  • CLL1 CAR of the present invention can also be “multi-chain CARs” as previously mentioned, which means that the extracellular binding domain and the signaling domains are preferably located on different polypeptide chains, whereas co-stimulatory domains may be located on the same or a third polypeptide.
  • Such multi-chain CARs can be derived from Fc ⁇ RI (Ravetch et al, 1989), by replacing the high affinity IgE binding domain of Fc ⁇ RI alpha chain by an extracellular ligand-binding domain such as scFv, whereas the N and/or C-termini tails of Fc ⁇ RI beta and/or gamma chains are fused to signal transducing domains and co-stimulatory domains respectively.
  • the extracellular ligand binding domain has the role of redirecting T-cell specificity towards cell targets, while the signal transducing domains activate or reduce the immune cell response.
  • the fact that the different polypeptides derive from the alpha, beta and gamma polypeptides from Fc ⁇ RI are transmembrane polypeptides sitting in juxtamembrane position provides a more flexible architecture to CARs, improving specificity towards the targeted molecule and reducing background activation of immune cells as described in WO2014/039523.
  • extracellular ligand-binding domain is defined as an oligo- or polypeptide that is capable of binding a ligand.
  • the domain will be capable of interacting with a cell surface molecule.
  • the extracellular ligand-binding domain may be chosen to recognize a ligand that acts as a cell surface marker on target cells associated with a particular disease state. It can be for instance binding domains derived from a ligand, a receptor, human or mice antibodies or antigen recognition domains derived from camels or cartilaginous fish.
  • said extracellular ligand-binding domain comprises a single chain antibody fragment (scFv) comprising the light (V L ) and the heavy (V H ) variable fragment of a target antigen specific monoclonal anti CLL1 antibody joined by a flexible linker.
  • scFv single chain antibody fragment
  • V L and V H are preferably selected from the antibodies referred to in the literature as SCO02-357, SCO2-378 and SC02-161 in WO2005/00894 (Applicant: Crucell Holland BV); M26, M31, G4, M22, M29, M2, M5, G12 in WO2013/169625 (Applicant: Cellerant Therapeutics); and 21.26, 1075.7 in WO2009/051974 (Applicant: Nuvelo Inc.), and more particularly as comprising CDRs from VH and VL domains of monoclonal anti-CLL1 antibodies selected from SEQ ID NO. 119 to 190 described below.
  • the CDR sequences of VH chain from monoclonal anti-CLL1 antibody may be chosen among GSISSSNWWS (SEQ ID NO 119), WIGEIYHSGSPDY (SEQ ID NO 120), KVSTGGFFDY (SEQ ID NO 121), and GSISSSNWWS (SEQ ID NO 122), WIGEIYHSGSPNY (SEQ ID NO 123), RSSSGGFFDY (SEQ ID NO 124), and GSISSSNWWS (SEQ ID NO 125), WIGEIYHSGSPNY (SEQ ID NO 126), RQTTAGSFDY (SEQ ID NO 127), and GYTFTSYFIH (SEQ ID NO 131), WIGFINPYNDGSKY (SEQ ID NO 132), TRDDGYYGYAMDY (SEQ ID NO 133), and GYTFTSYVMH (SEQ ID NO 137), WIGYINPYNDGTKY (SEQ ID NO 138), ARPIYFDNDY (SEQ ID NO 139),
  • the CDR sequences of VL chain from monoclonal anti-CLL1 antibody may be chosen among: QSISSYLN (SEQ ID NO 128), LLIYAASSLQS (SEQ ID NO 129), QQSYSTPP (SEQ ID NO 130), and QELSGYLS (SEQ ID NO 134), RLIYAASTLDS (SEQ ID NO 135), LQYAIYPY (SEQ ID NO 136), and ESVDSYGNSFMH (SEQ ID NO 140), LLIYLASNLES (SEQ ID NO 141), QQNNYDPW (SEQ ID NO 142), HDISNYLN (SEQ ID NO 146), LLIYYTSRLHS (SEQ ID NO 147), QQGKTLLW (SEQ ID NO 148), and QNLLNSGNQKKYLN (SEQ ID NO 152), LLIYWASTRES (SEQ ID NO 153), QNDYSYPF (SEQ ID NO 154), and QDINKYIA (SEQ ID NO
  • the extracellular domain and the transmembrane domain are preferably linked together by a flexible linker comprising the sequence SEQ ID NO.10.
  • said anti-CLL1 CARs preferentially comprise an extracellular ligand-biding domain comprising a polypeptide sequence displaying at least 90%, 95% 97% or 99% identity with an amino acid sequence selected from the group consisting of SEQ ID NO: 11 to SEQ ID NO: 34 (see Table 2).
  • recombinant antibody as used herein, is meant an antibody or antibody fragment which is generated using recombinant DNA technology, such as, for example, an antibody or antibody fragment expressed by a bacteriophage, a yeast expression system or a mammalian cell expression system.
  • the term should also be construed to mean an antibody or antibody fragment which has been generated by the synthesis of a DNA molecule encoding the antibody or antibody fragment and which DNA molecule expresses an antibody or antibody fragment protein, or an amino acid sequence specifying the antibody or antibody fragment, wherein the DNA or amino acid sequence has been obtained using recombinant or synthetic DNA or amino acid sequence technology which is available and well known in the art.
  • the present invention discloses a CLL1 specific single-chain chimeric antigen receptor (anti-CLL1 scCAR) as described above, wherein said extra cellular ligand binding-domain comprises VH and VL chains which are humanized.
  • the polypeptides include a humanized heavy chain variable region and a humanized light chain variable region.
  • the polypeptides may include the framework (FR) regions of the light and heavy chain variable regions of a human antibody, while retaining substantially the antigen-binding specificity of a parental monoclonal antibody.
  • the humanized heavy chain variable region and/or the humanized light chain variable region are at least about 87% humanized, at least about 90% humanized, at least about 95% humanized, at least about 98% humanized, or at least about 100% humanized, excluding the complementary-determining regions (CDRs).
  • the antigen-binding polypeptides molecules may be derived from monoclonal antibody donors (e.g., mouse monoclonal antibody donors) and may include CDRs from the monoclonal antibodies (e.g., mouse monoclonal CDRs).
  • monoclonal antibody antibody produced by a laboratory-grown cell clone, either of a hybridoma or a virus-transformed lymphocyte that is more abundant and uniform than natural antibody and is able to bind specifically to a single site on CLL1 antigen. They are monospecific antibodies that are made by identical immune cells that are all clones of a unique parent cell, in contrast to polyclonal antibodies which are made from several different immune cells. Monoclonal antibodies have monovalent affinity, in that they bind to the same epitope.
  • a humanized antibody can be produced using a variety of techniques known in the art, including but not limited to, CDR-grafting (see, e.g., European Patent No. EP 239,400; International Publication No. WO 91/09967; and U.S. Pat. Nos. 5,225,539, 5,530,101, and 5,585,089, each of which is incorporated herein in its entirety by reference), veneering or resurfacing (see, e.g., European Patent Nos.
  • framework substitutions are identified by methods well-known in the art, e.g., by modeling of the interactions of the CDR and framework residues to identify framework residues important for antigen binding and sequence comparison to identify unusual framework residues at particular positions. (See, e.g., Queen et al., U.S. Pat. No. 5,585,089; and Riechmann et al., 1988, Nature, 332:323, which are incorporated herein by reference in their entireties.).
  • Conservative amino acid substitutions are ones in which the amino acid residue is replaced with an amino acid residue having a similar side chain.
  • Families of amino acid residues having similar side chains have been defined in the art. These families include amino acids with basic side chains (e.g., lysine, arginine, histidine), acidic side chains (e.g., aspartic acid, glutamic acid), uncharged polar side chains (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine, tryptophan), nonpolar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine), beta-branched side chains (e.g., threonine, valine, isoleucine) and aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, histidine).
  • basic side chains
  • one or more amino acid residues within a anti-CLL1 CAR of the invention can be replaced with other amino acid residues from the same side chain family and the altered anti-CLL1 CAR can be tested for the ability to bind CLL1 using the functional assays described herein.
  • An anti-CLL1 CAR of the invention optionally includes a suicide domain, which is intended to deplete the immune cells endowed with the CAR in the event these later would cause adverse effects in vivo.
  • a suicide domain can be obtained, for instance, by including two copies of a CD20 mimotope, preferably of sequence CPYSNPSLCS (SEQ ID NO. 113), into the CAR polypeptide sequence. Said two copies of a CD20 mimotope can be linked to each other and also to the V L by a linker. They can also be inserted between the anti-CLL1 scFv and the hinge (such as CD8alpha), by using an optional linker.
  • the CD20 mimetopes can be bound by anti-CD20 antibodies, such as Rituximab (McLaughlin P, et al. 1998).
  • the anti-CLL1 CAR of the present invention may thus comprise a VH and a VL chains which are able to bind to CLL1 cell surface antigen, optionally humanized, a linker L, a suicide domain, a hinge or part of it, a transmembrane domain, a co-stimulatory domain and a stimulatory domain.
  • the present invention discloses an anti-CLL1 specific single-chain chimeric antigen receptor (“anti-CLL1 scCAR” or “scCAR”) having one of the polypeptide structure selected from V 1 to V6, as illustrated in FIG. 2 and Tables 3-8, said structure comprising an extra cellular ligand binding-domain comprising VH and VL from a monoclonal anti-CLL1 antibody, a hinge, a transmembrane domain, a cytoplasmic domain including a signaling domain and a co-stimulatory domain.
  • anti-CLL1 scCAR anti-CLL1 specific single-chain chimeric antigen receptor
  • the present invention discloses a CLL1 specific scCAR as described above, wherein said structure V1, V3 or V5 comprises a Fc ⁇ RIII ⁇ , CD8 alpha or IgG1 hinge and a CD8 alpha transmembrane domain.
  • said CLL1 specific scCAR comprises the co-stimulatory domain 4-1BB or the CD28, or more preferably the 4-1BB co-stimulatory domain.
  • the present invention discloses a CLL1 specific scCAR as described above, wherein said structure V1, V3 or V5 comprises a Fc ⁇ RIII ⁇ , CD8 alpha or IgG1 hinge and a 4-1BB transmembrane domain.
  • the present invention discloses a CLL1 specific scCAR as described above, wherein said structure V1, V3 or V5 comprises a Fc ⁇ RIII ⁇ , CD8 alpha or IgG1, a 4-1BB cytoplasmic domain and a CD8 alpha transmembrane domain.
  • the anti-CLL1 scCAR of the invention has one of the polypeptide structure selected from V1 to V6, as illustrated in FIG. 2 , said structure comprising an extra cellular ligand binding-domain comprising VH and VL from a monoclonal anti-CLL1 antibody, a hinge, a transmembrane domain, a cytoplasmic domain including a CD3 zeta signaling domain and a co-stimulatory domain, said CD3 zeta signaling domain preferably having a sequence SEQ ID NO.9.
  • the anti-CLL1 scCAR of the invention has one of the polypeptide structure selected from V1 to V6, as illustrated in FIG. 2 , said structure comprising an extra cellular ligand binding-domain comprising VH and VL from a monoclonal anti-CLL1 antibody, a hinge, a transmembrane domain, a cytoplasmic domain including a CD3 zeta signaling domain and a 4-1BB co-stimulatory domain, said 4-1BB co-stimulatory domain preferably having a sequence SEQ ID NO.8.
  • the present invention discloses anti-CLL1 scCAR having one of the polypeptide structure selected from V1 to V6, as illustrated in FIG. 2 , said structure comprising an extra cellular ligand binding-domain comprising VH and VL from a monoclonal anti-CLL1 antibody, a Fc ⁇ RIII ⁇ hinge, a Cd8 ⁇ transmembrane domain, preferably having SEQ ID NO.6, a cytoplasmic domain including a CD3 zeta signaling domain and a co-stimulatory domain.
  • the present invention discloses anti-CLL1 scCAR having one of the polypeptide structure selected from V1 to V6, as illustrated in FIG. 2 , said structure comprising an extra cellular ligand binding-domain comprising VH and VL from a monoclonal anti-CLL1 antibody, a Cd8 ⁇ hinge, a CD8 ⁇ transmembrane domain, preferably having SEQ ID NO.6, a cytoplasmic domain including a CD3 zeta signaling domain and a co-stimulatory domain.
  • the present invention discloses anti-CLL1 scCAR having one of the polypeptide structure selected from V1 to V6, as illustrated in FIG. 2 , said structure comprising an extra cellular ligand binding-domain comprising VH and VL from a monoclonal anti-CLL1 antibody, a IgG1 hinge, a CD8 ⁇ transmembrane domain, preferably having SEQ ID NO.6, a cytoplasmic domain including a CD3 zeta signaling domain and a co-stimulatory domain.
  • the present invention discloses anti-CLL1 scCAR having one of the polypeptide structure selected from V1 to V6, as illustrated in FIG. 2 , said structure comprising an extra cellular ligand binding-domain comprising VH and VL from a monoclonal anti-CLL1 antibody, a Fc ⁇ RIII ⁇ hinge, a 4-1BB transmembrane domain, preferably having SEQ ID NO.7, a cytoplasmic domain including a CD3 zeta signaling domain and a co-stimulatory domain.
  • the present invention discloses anti-CLL1 scCAR having one of the polypeptide structure selected from V1 to V6, as illustrated in FIG. 2 , said structure comprising an extra cellular ligand binding-domain comprising VH and VL from a monoclonal anti-CLL1 antibody, a Cd8 ⁇ hinge, a 4-1BB transmembrane domain, preferably having SEQ ID NO.7, a cytoplasmic domain including a CD3 zeta signaling domain and a co-stimulatory domain.
  • the present invention discloses anti-CLL1 scCAR having one of the polypeptide structure selected from V1 to V6, as illustrated in FIG. 2 , said structure comprising an extra cellular ligand binding-domain comprising VH and VL from a monoclonal anti-CLL1 antibody, a IgG1 hinge, a 4-1BB transmembrane domain, preferably having SEQ ID NO.7, a cytoplasmic domain including a CD3 zeta signaling domain and a co-stimulatory domain.
  • the present invention discloses an anti-CLL1 specific scCAR having a V1 polypeptide structure, as illustrated in FIG. 2 , said structure comprising an extra cellular ligand binding-domain comprising VH and VL from a monoclonal anti-CLL1 antibody, a Fc ⁇ RIII ⁇ hinge preferably with SEQ ID NO.3, a Cd8 ⁇ transmembrane domain, preferably with SEQ ID NO.6, a cytoplasmic domain including a CD3 zeta signaling domain, preferably with SEQ ID NO.9, and a 4-1BB co-stimulatory domain, preferably with SEQ ID NO.8.
  • the present invention discloses an anti-CLL1 specific scCAR having a V1 polypeptide structure, as illustrated in FIG. 2 , said structure comprising an extra cellular ligand binding-domain comprising VH and VL from a monoclonal anti-CLL1 antibody, a Fc ⁇ RIII ⁇ hinge preferably with SEQ ID NO.3, a Cd8 ⁇ transmembrane domain, preferably with SEQ ID NO.6, a cytoplasmic domain including a CD3 zeta signaling domain, preferably with SEQ ID NO.9, and a 4-1BB co-stimulatory domain, preferably with SEQ ID NO.8, wherein said VH chain having at least 80% identity with SEQ ID NO.11-13, 15, 17, 19, 21, 23, 25, 27, 29, 31 or33 and said VL having at least 80% identity with SEQ ID NO.14, 16, 18, 20, 22, 24, 26, 28, 30, 32 or 34.
  • the present invention discloses an anti-CLL1 specific scCAR having a V3 polypeptide structure, as illustrated in FIG. 2 , said structure comprising an extra cellular ligand binding-domain comprising VH and VL from a monoclonal anti-CLL1 antibody, a CD8 ⁇ hinge preferably with SEQ ID NO.4, a Cd8 ⁇ transmembrane domain, preferably with SEQ ID NO.6, a cytoplasmic domain including a CD3 zeta signaling domain, preferably with SEQ ID NO.9, and a 4-1BB co-stimulatory domain, preferably with SEQ ID NO.8.
  • the present invention discloses an anti-CLL1 specific scCAR having a V3 polypeptide structure, as illustrated in FIG. 2 , said structure comprising an extra cellular ligand binding-domain comprising VH and VL from a monoclonal anti-CLL1 antibody, a Cd8 ⁇ hinge preferably with SEQ ID NO.4, a Cd8 ⁇ transmembrane domain, preferably with SEQ ID NO.6, a cytoplasmic domain including a CD3 zeta signaling domain, preferably with SEQ ID NO.9, and a 4-1BB co-stimulatory domain, preferably with SEQ ID NO.8, wherein said VH chain having at least 80% identity with SEQ ID NO.11-13, 15, 17, 19, 21, 23, 25, 27, 29, 31 or 33 and said VL chain having at least 80% identity with SEQ ID NO.14, 16, 18, 20, 22, 24, 26, 28, 30, 32 or 34.
  • the present invention discloses an anti-CLL1 specific scCAR having a V5 polypeptide structure, as illustrated in FIG. 2 , said structure comprising an extra cellular ligand binding-domain comprising VH and VL from a monoclonal anti-CLL1 antibody, a IgG1 hinge preferably with SEQ ID NO.5, a Cd8 ⁇ transmembrane domain, preferably with SEQ ID NO.6, a cytoplasmic domain including a CD3 zeta signaling domain, preferably with SEQ ID NO.9, and a 4-1BB co-stimulatory domain, preferably with SEQ ID NO.8.
  • the present invention discloses an anti-CLL1 specific scCAR having a V5 polypeptide structure, as illustrated in FIG. 2 , said structure comprising an extra cellular ligand binding-domain comprising VH and VL from a monoclonal anti-CLL1 antibody, a IgG1 hinge preferably with SEQ ID NO.5, a Cd8 ⁇ transmembrane domain, preferably with SEQ ID NO.6, a cytoplasmic domain including a CD3 zeta signaling domain, preferably with SEQ ID NO.9, and a 4-1BB co-stimulatory domain, preferably with SEQ ID NO.8, wherein said VH chain having at least 80% identity with SEQ ID NO.11-13, 15, 17, 19, 21, 23, 25, 27, 29, 31 or 33 and said VL chain having at least 80% identity with SEQ ID NO.14, 16, 18, 20, 22, 24, 26, 28, 30, 32 or 34.
  • the present invention discloses an anti-CLL1 specific scCAR having a V1 polypeptide structure, as illustrated in FIG. 2 , said polypeptide having at least 80% identity with SEQ ID NO. 35, 41, 47, 53, 59, 65, 71, 77, 83, 89, 95, 101 or 107.
  • said anti-CLL1 specific scCAR having a V1 polypeptide structure, as illustrated in FIG. 2 , said structure comprising an extra cellular ligand binding-domain comprising VH and VL from a monoclonal anti-CLL1 antibody, a Fc ⁇ RIII ⁇ hinge preferably with SEQ ID NO.3, a CD8 ⁇ transmembrane domain, preferably with SEQ ID NO.6, a cytoplasmic domain including a CD3 zeta signaling domain, preferably with SEQ ID NO.9, and a 4-1BB co-stimulatory domain, preferably with SEQ ID NO.8, wherein said VH chain having at least 80% identity with SEQ ID NO.11,13, 15, 17, 19, 21, 23, 25, 27, 29, 31 or 33 and said VL having at least 80% identity with SEQ ID NO.14, 16, 18, 20, 22, 24, 26, 28, 30, 32 or 34, and wherein said polypeptide has at least 80% identity with SEQ ID NO. 35, 41, 47, 53, 59, 65,
  • the present invention discloses an anti-CLL1 specific scCAR of structure V3, as illustrated in FIG. 2 , said polypeptide having at least 80% identity with SEQ ID NO. 37, 43, 49, 55, 61, 67, 73, 79, 85, 91, 97, 103 or 109.
  • the present invention discloses an anti-CLL1 specific scCAR having a V3 polypeptide structure, as illustrated in FIG. 2 , said structure comprising an extra cellular ligand binding-domain comprising VH and VL from a monoclonal anti-CLL1 antibody, a Cd8 ⁇ hinge preferably with SEQ ID NO.4, a Cd8 ⁇ transmembrane domain, preferably with SEQ ID NO.6, a cytoplasmic domain including a CD3 zeta signaling domain, preferably with SEQ ID NO.9, and a 4-1BB co-stimulatory domain, preferably with SEQ ID NO.8, wherein said VH chain having at least 80% identity with SEQ ID NO.11,13, 15, 17, 19, 21, 23, 25, 27, 29, 31 or 33 and said VL chain having at least 80% identity with SEQ ID NO.14, 16, 18, 20, 22, 24, 26, 28, 30, 32 or 34, and wherein said polypeptide has at least 80% identity with SEQ ID NO. 37, 43, 49, 55,
  • the present invention discloses an anti-CLL1 specific scCAR of structure V5, as illustrated in FIG. 2 , said polypeptide having at least 80% identity with SEQ ID NO.39, 45, 51, 57, 63, 69, 75, 81, 87, 93, 99, 105 or 111.
  • the present invention discloses an anti-CLL1 specific scCAR having a V5 polypeptide structure, as illustrated in FIG. 2 , said structure comprising an extra cellular ligand binding-domain comprising VH and VL from a monoclonal anti-CLL1 antibody, a IgG1 hinge preferably with SEQ ID NO.5, a Cd8 ⁇ transmembrane domain, preferably with SEQ ID NO.6, a cytoplasmic domain including a CD3 zeta signaling domain, preferably with SEQ ID NO.9, and a 4-1BB co-stimulatory domain, preferably with SEQ ID NO.8, wherein said VH chain having at least 80% identity with SEQ ID NO.11,13, 15, 17, 19, 21, 23, 25, 27, 29, 31 or 33 and said VL chain having at least 80% identity with SEQ ID NO.14, 16, 18, 20, 22, 24, 26, 28, 30, 32 or 34, and wherein said polypeptide having at least 80% identity with SEQ ID NO.39, 45, 51, 57,
  • the present invention more particularly discloses a CLL1 single-chain specific chimeric antigen receptor (scCAR) having a polypeptide structure V1, V3 or V5 as illustrated in FIG. 2 , and described above said structure comprising an extra cellular ligand binding-domain VH from a monoclonal anti-CLL1 antibody comprising the following CDR sequences: GSISSSNWWS (SEQ ID NO 119), WIGEIYHSGSPDY (SEQ ID NO 120), KVSTGGFFDY (SEQ ID NO 121), and GSISSSNWWS (SEQ ID NO 122), WIGEIYHSGSPNY (SEQ ID NO 123), RSSSGGFFDY (SEQ ID NO 124), and GSISSSNWWS (SEQ ID NO 125), WIGEIYHSGSPNY (SEQ ID NO 126), RQTTAGSFDY (SEQ ID NO 127), and GYTFTSYFIH (SEQ ID NO 131), WIGFINPYNDGSKY (SEQ ID NO
  • an extra cellular ligand binding-domain VL from a monoclonal anti-CLL1 antibody comprising the following CDR sequences: QSISSYLN (SEQ ID NO 128), LLIYAASSLQS (SEQ ID NO 129), QQSYSTPP (SEQ ID NO 130), and QELSGYLS (SEQ ID NO 134), RLIYAASTLDS (SEQ ID NO 135), LQYAIYPY (SEQ ID NO 136), and ESVDSYGNSFMH (SEQ ID NO 140), LLIYLASNLES (SEQ ID NO 141), QQNNYDPW (SEQ ID NO 142), HDISNYLN (SEQ ID NO 146), LLIYYTSRLHS (SEQ ID NO 147), QQGKTLLW (SEQ ID NO 148), and QNLLNSGNQKKYLN (SEQ ID NO 152), LLIYWASTRES (SEQ ID NO 153), QNDYSYPF (SEQ ID NO 154), and QDIN
  • a hinge a transmembrane domain and a cytoplasmic domain including a CD3 zeta signaling domain and a co-stimulatory domain from 4-1BB.
  • the present invention discloses an anti-CLL1 single-chain specific chimeric antigen receptor (anti-CLL1 scCAR) as above, wherein said extra cellular ligand binding-domain VH and VL is humanized.
  • the present invention discloses a CLL1 single-chain specific chimeric antigen receptor (scCAR) as described above, wherein said extra cellular ligand binding-domain VH from a monoclonal anti-CLL1 antibody comprise at least one of the following sequences:
  • the present invention discloses a CLL1 single-chain specific chimeric antigen receptor (scCAR) as described above, wherein said extra cellular ligand binding-domain VH from a monoclonal anti-CLL1 antibody comprise at least one of the following sequences:
  • VL from a monoclonal anti-CLL1 antibody comprise at least one of the following sequences:
  • the present invention also discloses a CLL1 specific scCAR as previously defined, further comprising another extracellular ligand binding domain which is not specific for CLL1, such as CD33 antigen, CD44 antigen, CD47 antigen, CD123 antigen, CD96 antigen and T-cell immunoglobulin mucin-3 (TIM-3).
  • a CLL1 specific scCAR as previously defined, further comprising another extracellular ligand binding domain which is not specific for CLL1, such as CD33 antigen, CD44 antigen, CD47 antigen, CD123 antigen, CD96 antigen and T-cell immunoglobulin mucin-3 (TIM-3).
  • the present invention discloses a CLL1 specific scCAR as above, further comprising a signal peptide, preferably of SEQ ID NO 1 or SEQ ID NO 2, in order to help the CAR polypeptide to reach the immune cell's membrane.
  • the present invention discloses a CLL1 specific scCAR as above, wherein a linker of SEQ ID NO 10 is inserted between VH and VL.
  • the present invention also relates to polynucleotides and vectors allowing heterologous expression into cells of the anti-CLL1 CAR according to the invention, encoding the polypeptides sequences which have been previously detailed.
  • the polynucleotides may be included in an expression cassette or expression vector (e.g. a plasmid for introduction into a bacterial host cell, or a viral vector such as a baculovirus vector for transfection of an insect host cell, or a plasmid or viral vector such as a lentivirus for transfection of a mammalian host cell).
  • an expression cassette or expression vector e.g. a plasmid for introduction into a bacterial host cell, or a viral vector such as a baculovirus vector for transfection of an insect host cell, or a plasmid or viral vector such as a lentivirus for transfection of a mammalian host cell.
  • the different nucleic acid sequences can be included in one polynucleotide or vector which comprises a nucleic acid sequence encoding ribosomal skip sequence such as a sequence encoding a 2A peptide.
  • 2A peptides which were identified in the Aphthovirus subgroup of picornaviruses, causes a ribosomal “skip” from one codon to the next without the formation of a peptide bond between the two amino acids encoded by the codons (see (Donnelly and Elliott 2001; Atkins, Wills et al. 2007; Doronina, Wu et al. 2008)).
  • cognate is meant three nucleotides on an mRNA (or on the sense strand of a DNA molecule) that are translated by a ribosome into one amino acid residue.
  • two polypeptides can be synthesized from a single, contiguous open reading frame within an mRNA when the polypeptides are separated by a 2A oligopeptide sequence that is in frame.
  • Such ribosomal skip mechanisms are well known in the art and are known to be used by several vectors for the expression of several proteins encoded by a single messenger RNA.
  • a secretory signal sequence (also known as a leader sequence, prepro sequence or pre sequence) is provided in polynucleotide sequence or vector sequence.
  • the secretory signal sequence is operably linked to the transmembrane nucleic acid sequence, i.e., the two sequences are joined in the correct reading frame and positioned to direct the newly synthesized polypeptide into the secretory pathway of the host cell.
  • Secretory signal sequences are commonly positioned 5′ to the nucleic acid sequence encoding the polypeptide of interest, although certain secretory signal sequences may be positioned elsewhere in the nucleic acid sequence of interest (see, e.g., Welch et al., U.S. Pat. No. 5,037,743; Holland et al., U.S. Pat. No. 5,143,830).
  • the signal peptide comprises the amino acid sequence SEQ ID NO: 1 and 2 or at least 90%, 95% 97% or 99% sequence identity with SEQ ID NO: 1 and/or 2.
  • the nucleic acid sequences of the present invention are codon-optimized for expression in mammalian cells, preferably for expression in human cells. Codon-optimization refers to the exchange in a sequence of interest of codons that are generally rare in highly expressed genes of a given species by codons that are generally frequent in highly expressed genes of such species, such codons encoding the amino acids as the codons that are being exchanged.
  • the present invention encompasses the different means to express the anti-CLL1 Chimeric Antigen Receptor (CAR) described herein in immune cells
  • Methods for introducing a polynucleotide construct into cells include as non-limiting examples stable transformation methods wherein the polynucleotide construct encoding said CAR is integrated into the genome of the cell, transient transformation methods wherein the polynucleotide construct is not integrated into the genome of the cell and virus mediated methods.
  • Said polynucleotides may be introduced into a cell by for example, recombinant viral vectors (e.g. retroviruses, adenoviruses), liposome and the like.
  • transient transformation methods include for example microinjection, electroporation or particle bombardment, cell fusion.
  • Said polynucleotides may be included in vectors, more particularly plasmids or virus, in view of being expressed in cells.
  • Said plasmid vector can comprise a selection marker which provides for identification and/or selection of cells which received said vector.
  • Said vector can comprise a nucleic acid sequence encoding ribosomal skip sequence such as a sequence encoding a 2A peptide.
  • 2A peptides which were identified in the Aphthovirus subgroup of picornaviruses, causes a ribosomal “skip” from one codon to the next without the formation of a peptide bond between the two amino acids encoded by the codons (see Donnelly et al., J. of General Virology 82: 1013-1025 (2001); Donnelly et al., J. of Gen. Virology 78: 13-21 (1997); Doronina et al., Mol. And. Cell. Biology 28(13): 4227-4239 (2008); Atkins et al., RNA 13: 803-810 (2007)).
  • cognate is meant three nucleotides on an mRNA (or on the sense strand of a DNA molecule) that are translated by a ribosome into one amino acid residue.
  • two polypeptides can be synthesized from a single, contiguous open reading frame within an mRNA when the polypeptides are separated by a 2A oligopeptide sequence that is in frame.
  • Such ribosomal skip mechanisms are well known in the art and are known to be used by several vectors for the expression of several proteins encoded by a single messenger RNA.
  • polynucleotides encoding polypeptides according to the present invention can be mRNA which is introduced directly into the cells, for example by electroporation.
  • the inventors determined the optimal condition for mRNA electroporation in T-cell.
  • the inventor used the cytoPulse technology which allows, by the use of pulsed electric fields, to transiently permeabilize living cells for delivery of material into the cells.
  • the technology based on the use of PulseAgile® (BTX Havard Apparatus, 84 October Hill Road, Holliston, Mass. 01746, USA) electroporation waveforms grants the precise control of pulse duration, intensity as well as the interval between pulses (U.S. Pat. No.
  • the first high electric field pulses allow pore formation, while subsequent lower electric field pulses allow moving the polynucleotide into the cell.
  • scCAR can be introduced as transgenes encoded by one plasmid vector.
  • Said plasmid vector can also contain a selection marker which provides for identification and/or selection of cells which received said vector.
  • Polypeptides may be synthesized in situ in the cell as a result of the introduction of polynucleotides encoding said polypeptides into the cell. Alternatively, said polypeptides could be produced outside the cell and then introduced thereto.
  • Methods for introducing a polynucleotide construct into cells are known in the art and including as non limiting examples stable transformation methods wherein the polynucleotide construct is integrated into the genome of the cell, transient transformation methods wherein the polynucleotide construct is not integrated into the genome of the cell and virus mediated methods.
  • Said polynucleotides may be introduced into a cell by for example, recombinant viral vectors (e.g.
  • retroviruses adenoviruses
  • liposome adenoviruses
  • transient transformation methods include for example microinjection, electroporation or particle bombardment.
  • Said polynucleotides may be included in vectors, more particularly plasmids or virus, in view of being expressed in cells.
  • the immune cells, particularly T-cells of the present invention can be further activated and expanded generally using methods as described, for example, in U.S. Pat. Nos. 6,352,694; 6,534,055; 6,905,680; 6,692,964; 5,858,358; 6,887,466; 6,905,681; 7,144,575; 7,067,318; 7,172,869; 7,232,566; 7,175,843; 5,883,223; 6,905,874; 6,797,514; 6,867,041; and U.S. Patent Application Publication No. 20060121005.
  • T cells can be expanded in vitro or in vivo.
  • the T cells of the invention are expanded by contact with an agent that stimulates a CD3 TCR complex and a co-stimulatory molecule on the surface of the T cells to create an activation signal for the T-cell.
  • an agent that stimulates a CD3 TCR complex and a co-stimulatory molecule on the surface of the T cells to create an activation signal for the T-cell.
  • chemicals such as calcium ionophore A23187, phorbol 12-myristate 13-acetate (PMA), or mitogenic lectins like phytohemagglutinin (PHA) can be used to create an activation signal for the T-cell.
  • T cell populations may be stimulated in vitro such as by contact with an anti-CD3 antibody, or antigen-binding fragment thereof, or an anti-CD2 antibody immobilized on a surface, or by contact with a protein kinase C activator (e.g., bryostatin) in conjunction with a calcium ionophore.
  • a protein kinase C activator e.g., bryostatin
  • a ligand that binds the accessory molecule is used for co-stimulation of an accessory molecule on the surface of the T cells.
  • a population of T cells can be contacted with an anti-CD3 antibody and an anti-CD28 antibody, under conditions appropriate for stimulating proliferation of the T cells.
  • Conditions appropriate for T cell culture include an appropriate media (e.g., Minimal Essential Media or RPMI Media 1640 or, X-vivo 5, (LonzaLONZA®)) that may contain factors necessary for proliferation and viability, including serum (e.g., fetal bovine or human serum), interleukin-2 (IL-2), insulin, IFN-g, ll-4, ll-7, GM-CSF, -10, -2, ll-15, TGFp, and TNF- or any other additives for the growth of cells known to the skilled artisan.
  • Other additives for the growth of cells include, but are not limited to, surfactant, plasmanate, and reducing agents such as N-acetylcysteine and 2-mercaptoethanoi.
  • Media can include RPMI1640, A1M-V, DMEM, MEM, a-MEM, F-12, X-Vivo 1, and X-Vivo 20, Optimizer, with added amino acids, sodium pyruvate, and vitamins, either serum-free or supplemented with an appropriate amount of serum (or plasma) or a defined set of hormones, and/or an amount of cytokine(s) sufficient for the growth and expansion of T cells.
  • Antibiotics e.g., penicillin and streptomycin, are included only in experimental cultures, not in cultures of cells that are to be infused into a subject.
  • the target cells are maintained under conditions necessary to support growth, for example, an appropriate temperature (e.g., 37° C.) and atmosphere (e.g., air plus 5% CO 2 ). T cells that have been exposed to varied stimulation times may exhibit different characteristics.
  • said cells can be expanded by co-culturing with tissue or cells. Said cells can also be expanded in vivo, for example in the subject's blood after administrating said cell into the subject.
  • a “Cell” according to the present invention generally refers to a cell of hematopoietic origin functionally involved in the initiation and/or execution of innate and/or adaptative immune response.
  • Cell according to the present invention is preferably an isolated immune cell, and more preferably a T-cell obtained from a donor. Said immune cell according to the present invention can also be derived from a stem cell.
  • the stem cells can be adult stem cells, non-human embryonic stem cells, more particularly non-human stem cells, cord blood stem cells, progenitor cells, bone marrow stem cells, induced pluripotent stem cells, totipotent stem cells or hematopoietic stem cells.
  • Representative human cells are CD34+ cells.
  • Said isolated cell can also be a dendritic cell, killer dendritic cell, a mast cell, a NK-cell, a B-cell or a T-cell selected from the group consisting of inflammatory T-lymphocytes, cytotoxic T-lymphocytes, regulatory T-lymphocytes or helper T-lymphocytes.
  • said cell can be derived from the group consisting of CD4+ T-lymphocytes and CD8+ T-lymphocytes.
  • a source of cells can be obtained from a subject through a variety of non-limiting methods.
  • Cells can be obtained from a number of non-limiting sources, including peripheral blood mononuclear cells, bone marrow, lymph node tissue, cord blood, thymus tissue, tissue from a site of infection, ascites, pleural effusion, spleen tissue, and tumors.
  • any number of T cell lines available and known to those skilled in the art may be used.
  • said cell can be derived from a healthy donor, from a patient diagnosed with cancer or from a patient diagnosed with an infection.
  • said cell is part of a mixed population of cells which present different phenotypic characteristics.
  • a cell line obtained from a transformed T-cell according to the method previously described Modified cells resistant to an immunosuppressive treatment and susceptible to be obtained by the previous method are encompassed in the scope of the present invention.
  • the present invention provides T-cells or a population of primary T-cells, endowed with a CLL1 CAR as described above, that do not express functional TCR and that a reactive towards CLL1 positive cells, for their allogeneic transplantation into patients.
  • the present invention provides T-cells or a population of T-cells endowed with a CLL1 scCAR and that a reactive towards CLL1 positive cells as described above, that do not express a functional TCR and are resistant to a selected drug, for their allogeneic transplantation into patients treated with said selected drug.
  • the present invention encompasses the method of preparing engineered immune cells for immunotherapy comprising introducing ex-vivo into said immune cells the polynucleotides or vectors encoding the CLL1 CAR according to transformation methods as previously described in WO2014/130635WO2013176916, WO2013176915 and incorporated herein by reference.
  • said polynucleotides are introduced into the immune cells by means of retroviral vectors in view of being stably integrated into the cell genome.
  • the present invention also aims to produce immune cells endowed with anti CLL1 CAR, which are less or non-alloreactive, which can be used in allogeneic treatments (i.e. with reduced risk of inducing Graft versus host reaction) and/or made resistant to various standard of care treatments).
  • said methods may further comprise the step of genetically modifying said immune cell by using at least one endonuclease.
  • exonuclease refers to any wild-type or variant enzyme capable of catalyzing the hydrolysis (cleavage) of bonds between nucleic acids within a DNA or RNA molecule, preferably a DNA molecule. Endonucleases do not cleave the DNA or RNA molecule irrespective of its sequence, but recognize and cleave the DNA or RNA molecule at specific polynucleotide sequences, further referred to as “target sequences” or “target sites”. Endonucleases can be classified as rare-cutting endonucleases when having typically a polynucleotide recognition site greater than 12 base pairs (bp) in length, more preferably of 14-55 bp.
  • Rare-cutting endonucleases can for example be a homing endonuclease (Paques and Duchateau 2007), a chimeric Zinc-Finger nuclease (ZFN) resulting from the fusion of engineered zinc-finger domains with the catalytic domain of a restriction enzyme such as FokI (Porteus and Carroll 2005), a TALE-nuclease, a Cas9 endonuclease from CRISPR system as described below (Gasiunas, Barrangou et al. 2012; Jinek, Chylinski et al.
  • a chemical or peptidic cleaver is conjugated either to a polymer of nucleic acids or to another DNA recognizing a specific target sequence, thereby targeting the cleavage activity to a specific sequence.
  • Chemical endonucleases also encompass synthetic nucleases like conjugates of orthophenanthroline, a DNA cleaving molecule, and triplex-forming oligonucleotides (TFOs), known to bind specific DNA sequences (Kalish and Glazer 2005).
  • Rare-cutting endonucleases can be used for inactivating genes at a locus or to integrate transgenes by homologous recombination (HR) i.e. by inducing DNA double-strand breaks (DSBs) at a locus and insertion of exogeneous DNA at this locus by gene repair mechanism (Perrin, Buckle et al. 1993; Rouet, Smih et al. 1994; Choulika, Perrin et al. 1995; Pingoud and Silva 2007).
  • HR homologous recombination
  • DSBs DNA double-strand breaks
  • TALE-nuclease TALEN
  • TALEN Transcription Activator Like Effector
  • the catalytic domain is preferably a nuclease domain and more preferably a domain having endonuclease activity, like for instance 1-Tevl, CoiE7, NucA and Fok-1.
  • the TALE domain can be fused to a meganuclease like for instance 1-Crel and 1-0nul or functional variant thereof.
  • said nuclease is a monomeric TALE-Nuclease.
  • a monomeric TALE-Nuclease is a TALE-Nuclease that does not require dimerization for specific recognition and cleavage, such as the fusions of engineered TAL repeats with the catalytic domain of 1-Tevl described in WO2012138927.
  • Transcription Activator like Effector are proteins from the bacterial species Xanthomonas comprise a plurality of repeated sequences, each repeat comprising di-residues in position 12 and 13 (RVD) that are specific to each nucleotide base of the nucleic acid targeted sequence.
  • Binding domains with similar modular base-per-base nucleic acid binding properties can also be derived from new modular proteins recently discovered by the applicant in a different bacterial species.
  • the new modular proteins have the advantage of displaying more sequence variability than TAL repeats.
  • RVDs associated with recognition of the different nucleotides are HD for recognizing C, NG for recognizing T, NI for recognizing A, NN for recognizing G or A, NS for recognizing A, C, G or T, HG for recognizing T, IG for recognizing T, NK for recognizing G, HA for recognizing C, ND for recognizing C, HI for recognizing C, HN for recognizing G, NA for recognizing G, SN for recognizing G or A and YG for recognizing T, TL for recognizing A, VT for recognizing A or G and SW for recognizing A.
  • critical amino acids 12 and 13 can be mutated towards other amino acid residues in order to modulate their specificity towards nucleotides A, T, C and G and in particular to enhance this specificity.
  • TALE-nuclease have been already described and used to stimulate gene targeting and gene modifications (Bach, Scholze et al. 2009; Moscou and Bogdanove 2009; Christian, Cermak et al. 2010; Li, Huang et al. 2011).
  • Engineered TAL-nucleases are available under the trade name TALENTM (CELLECTIS®, 8 rue de Ia Croix Jarry, 75013 Paris, France) and can be ordered from manufacturers, such as Life Technologies (Carlsbad, Calif., USA).
  • Preferred TALE-nucleases recognizing and cleaving the target sequence are described in PCT/EP2014/075317.
  • additional catalytic domain can be further introduced into the cell with said rare-cutting endonuclease to increase mutagenesis in order to enhance their capacity to inactivate targeted genes.
  • said additional catalytic domain is a DNA end processing enzyme.
  • DNA end-processing enzymes include 5-3′ exonucleases, 3-5′ exonucleases, 5-3′ alkaline exonucleases, 5′ flap endonucleases, helicases, hosphatase, hydrolases and template-independent DNA polymerases.
  • Non limiting examples of such catalytic domain comprise of a protein domain or catalytically active derivate of the protein domain selected from the group consisting of hExoI (EXO1_HUMAN), Yeast ExoI (EXO1_YEAST), E. coli ExoI, Human TREX2, Mouse TREX1, Human TREX1, Bovine TREX1, Rat TREX1, TdT (terminal deoxynucleotidyl transferase) Human DNA2, Yeast DNA2 (DNA2_YEAST).
  • EXO1_HUMAN hExoI
  • EXO1_YEAST Yeast ExoI
  • E. coli ExoI E. coli ExoI
  • Human TREX2 Mouse TREX1, Human TREX1, Bovine TREX1, Rat TREX1, TdT (terminal deoxynucleotidyl transferase) Human DNA2, Yeast DNA2 (DNA2_YEAST).
  • said additional catalytic domain has a 3′-5′-exonuclease activity, and in a more preferred embodiment, said additional catalytic domain is TREX, more preferably TREX2 catalytic domain (WO2012/058458). In another preferred embodiment, said catalytic domain is encoded by a single chain TREX2 polypeptide. Said additional catalytic domain may be fused to a nuclease fusion protein or chimeric protein according to the invention optionally by a peptide linker.
  • Cas9 endonuclease any genome engineering tool developed based on the RNA-guided Cas9 nuclease (Gasiunas, Barrangou et al. 2012; Jinek, Chylinski et al. 2012; Cong, Ran et al. 2013; Mali, Yang et al. 2013) from the type II prokaryotic CRISPR (Clustered Regularly Interspaced Short palindromic Repeats) adaptive immune system (see for review (Sorek, Lawrence et al. 2013)).
  • CRISPR Associated (Cas) system was first discovered in bacteria and functions as a defense against foreign DNA, either viral or plasmid.
  • CRISPR-mediated genome engineering first proceeds by the selection of target sequence often flanked by a short sequence motif, referred as the proto-spacer adjacent motif (PAM). Following target sequence selection, a specific crRNA, complementary to this target sequence is engineered. Trans-activating crRNA (tracrRNA) required in the CRISPR type II systems paired to the crRNA and bound to the provided Cas9 protein.
  • tracrRNA Trans-activating crRNA required in the CRISPR type II systems paired to the crRNA and bound to the provided Cas9 protein.
  • Cas9 acts as a molecular anchor facilitating the base pairing of tracRNA with cRNA (Deltcheva, Chylinski et al. 2011).
  • the dual tracrRNA:crRNA structure acts as guide RNA that directs the endonuclease Cas9 to the cognate target sequence.
  • Target recognition by the Cas9-tracrRNA:crRNA complex is initiated by scanning the target sequence for homology between the target sequence and the crRNA.
  • DNA targeting requires the presence of a short motif adjacent to the protospacer (protospacer adjacent motif—PAM).
  • PAM protospacer adjacent motif
  • TCR T-Cell Receptor
  • T-cell endowed with anti-CLL1 CAR of the present invention can be made less alloreactive, for instance, by inactivating at least one gene expressing one or more component of T-cell receptor (TCR) as described in WO 2013/176915.
  • TCR T-cell receptor
  • This inactivation can be combined with that of another gene, such as of a gene encoding or regulating HLA or ⁇ 2m protein expression. Accordingly, the risk of graft versus host syndrome and graft rejection is significantly reduced.
  • Methods of making cells less allogenic can comprise the step of inactivating at least one gene encoding a T-Cell Receptor (TCR) component, in particular TCRalpha and/or TCRbeta genes.
  • TCR T-Cell Receptor
  • the present invention encompasses an anti-CLL1 CAR expressing immune cell wherein at least one gene expressing one or more component of T-cell receptor (TCR) has been inactivated.
  • the present invention provides an anti-CLL1 CAR expressing T cell wherein at least one gene expressing one or more component of T-cell receptor (TCR) is inactivated.
  • the genetic modification of the method relies on the expression, in provided cells to engineer, of one rare-cutting endonuclease such that said rare-cutting endonuclease specifically catalyzes cleavage in one targeted gene thereby inactivating said targeted gene.
  • the nucleic acid strand breaks caused by the rare-cutting endonuclease are commonly repaired through the distinct mechanisms of homologous recombination or non-homologous end joining (NHEJ).
  • NHEJ non-homologous end joining
  • NHEJ non-homologous end joining
  • the step of inactivating at least a gene encoding a component of the T-cell receptor (TCR) into the cells of each individual sample comprises introducing into the cell a rare-cutting endonuclease able to disrupt at least one gene encoding a component of the T-cell receptor (TCR).
  • said cells of each individual sample are transformed with nucleic acid encoding a rare-cutting endonuclease capable of disrupting at least one gene encoding a component of the T-cell receptor (TCR), and said rare-cutting endonuclease is expressed into said cells.
  • said method of further engineer the immune cells involves introducing into said T cells polynucleotides, in particular mRNAs, encoding specific rare-cutting endonuclease to selectively inactivate the genes mentioned above by DNA cleavage.
  • said rare-cutting endonucleases are TALE-nucleases or Cas9 endonuclease.
  • TAL-nucleases have so far proven higher specificity and cleavage efficiency over the other types of rare-cutting endonucleases, making them the endonucleases of choice for producing of the engineered immune cells on a large scale with a constant turn-over.
  • anti-CLL1 CAR immune cells with one or more component of T-cell receptor (TCR) inactivated are intended to be used as a medicament.
  • anti-CLL1 CAR expressing immune cells of the invention can be further genetically engineered to make them resistant to immunosuppressive drugs or chemotherapy treatments, which are used as standard care for treating cancer associated with CLL1 positive malignant cell, especially AML.
  • cytotoxic agents such as anti-cancer drug
  • alkylating agents such as alkylating agents, anthracyclines, DNA methyltransferase inhibitors, platinum compounds and spindle poisons
  • novel therapies such as immunotherapies
  • chemotherapy agents can be detrimental to the establishment of robust anti-tumor immunocompetent cells due to the agents' non-specific toxicity profiles.
  • Small molecule-based therapies targeting cell proliferation pathways may also hamper the establishment of anti-tumor immunity. If chemotherapy regimens that are transiently effective can be combined with novel immunocompetent cell therapies then significant improvement in anti-neoplastic therapy might be achieved (for review (Dasgupta, McCarty et al. 2011).
  • drug resistance is conferred to said allogeneic cells to protect them from the toxic side-effects of chemotherapy agents.
  • the drug resistance of immune cells also permits their enrichment in or ex vivo, as T-cells which express the drug resistance gene will survive and multiply relative to drug sensitive cells.
  • the present invention relates to a method of engineering allogeneic cells suitable for immunotherapy wherein at least one gene encoding a T-cell receptor (TCR) component is inactivated and one gene is modified to confer drug resistance comprising:
  • the present invention relates to a method comprising:
  • the present invention also relates to a method of engineering allogeneic cells suitable for immunotherapy wherein at least one gene encoding a T-cell receptor (TCR) component is inactivated and one gene is modified to confer drug resistance comprising:
  • the present invention relates to a method comprising:
  • said drug resistance can be conferred to the T-cell by the expression of at least one drug resistance gene.
  • Said drug resistance gene refers to a nucleic acid sequence that encodes “resistance” to an agent, such as a chemotherapeutic agent (e.g. methotrexate).
  • a chemotherapeutic agent e.g. methotrexate
  • the expression of the drug resistance gene in a cell permits proliferation of the cells in the presence of the agent to a greater extent than the proliferation of a corresponding cell without the drug resistance gene.
  • the expression of the drug resistance gene in a cell permits proliferation of the cells in the presence of the agent and does not affect its activity.
  • a drug resistance gene of the invention can encode resistance to anti-metabolite, methotrexate, vinblastine, cisplatin, alkylating agents, anthracyclines, cytotoxic antibiotics, anti-immunophilins, their analogs or derivatives, and the like.
  • a drug resistance gene of the invention can confer resistance to a drug (or an agent), in particular an anti-cancer drug selected from aracytine, cytosine arabinoside, amsacrine, daunorubicine, idarubicine, novantrone, mitoxantrone, vepeside, etoposide (VP16), arsenic trioxyde, transretinoic acid, combination of arsenic trioxyde, transretinoic acid, mechlorethamine, procarbazine, chlorambucil, cytarabine, anthracyclines, 6-thioguanine, hydroxyurea, prednisone, and combination thereof.
  • a drug or an agent
  • an anti-cancer drug selected from aracytine, cytosine arabinoside, amsacrine, daunorubicine, idarubicine, novantrone, mitoxantrone, vepeside, etoposide (VP16), arsenic trioxyde
  • DHFR Dihydrofolate reductase
  • DHFR is an enzyme involved in regulating the amount of tetrahydrofolate in the cell and is essential to DNA synthesis.
  • Folate analogs such as methotrexate (MTX) inhibit DHFR and are thus used as anti-neoplastic agents in clinic.
  • MTX methotrexate
  • the drug resistance gene according to the present invention can be a nucleic acid sequence encoding a mutant form of human wild type DHFR (GenBankTM: AAH71996.1) which comprises at least one mutation conferring resistance to an anti-folate treatment, such as methotrexate.
  • mutant form of DHFR comprises at least one mutated amino acid at position G15, L22, F31 or F34, preferably at positions L22 or F31 (Schweitzer, Dicker et al. 1990); International application WO94/24277; US patent U.S. Pat. No. 6,642,043).
  • said DHFR mutant form comprises two mutated amino acids at position L22 and F31.
  • the serine residue at position 15 is preferably replaced with a tryptophan residue.
  • the leucine residue at position 22 is preferably replaced with an amino acid which will disrupt binding of the mutant DHFR to antifolates, preferably with uncharged amino acid residues such as phenylalanine or tyrosine.
  • the phenylalanine residue at positions 31 or 34 is preferably replaced with a small hydrophilic amino acid such as alanine, serine or glycine.
  • antifolate agent refers to a molecule directed to interfere with the folate metabolic pathway at some level.
  • antifolate agents include, e.g., methotrexate (MTX); aminopterin; trimetrexate (NeutrexinTM); edatrexate; N10-propargyl-5,8-dideazafolic acid (CB3717); ZD1694 (Tumodex), 5,8-dideazaisofolic acid (IAHQ); 5,10-dideazatetrahydrofolic acid (DDATHF); 5-deazafolic acid; PT523 (N alpha-(4-amino-4-deoxypteroyl)-N delta-hemiphthaloyl-L-ornithine); 10-ethyl-10-deazaaminopterin (DDATHF, lomatrexol); piritrexim; 10-EDAM; ZD1694; GW1843; Pemetrexate and
  • IMPDH2 ionisine-5′-monophosphate dehydrogenase II
  • the mutant or modified form of IMPDH2 is an IMPDH inhibitor resistance gene.
  • IMPDH inhibitors can be mycophenolic acid (MPA) or its prodrug mycophenolate mofetil (MMF).
  • MMF prodrug mycophenolate mofetil
  • the mutant IMPDH2 can comprises at least one, preferably two mutations in the MAP binding site of the wild type human IMPDH2 (NP_000875.2) that lead to a significantly increased resistance to IMPDH inhibitor.
  • the mutations are preferably at positions T333 and/or S351 (Yam, Jensen et al. 2006; Sangiolo, Lesnikova et al. 2007; Jonnalagadda, Brown et al. 2013).
  • the threonine residue at position 333 is replaced with an isoleucine residue and the serine residue at position 351 is replaced with a tyrosine residue.
  • Correspondence of amino acid positions described herein is frequently expressed in terms of the positions of the amino acids of the form of wild-type human IMPDH2 polypeptide set forth in NP_000875.2.
  • Calcineurin an ubiquitously expressed serine/threonine protein phosphatase that is involved in many biological processes and which is central to T-cell activation. Calcineurin is a heterodimer composed of a catalytic subunit (CnA; three isoforms) and a regulatory subunit (CnB; two isoforms). After engagement of the T-cell receptor, calcineurin dephosphorylates the transcription factor NFAT, allowing it to translocate to the nucleus and active key target gene such as IL2.
  • CnA catalytic subunit
  • CnB regulatory subunit
  • the drug resistance gene of the present invention can be a nucleic acid sequence encoding a mutant form of calcineurin resistant to calcineurin inhibitor such as FK506 and/or CsA.
  • said mutant form can comprise at least one mutated amino acid of the wild type calcineurin heterodimer a at positions: V314, Y341, M347, T351, W352, L354, K360, preferably double mutations at positions T351 and L354 or V314 and Y341.
  • the valine residue at position 341 can be replaced with a lysine or an arginine residue
  • the tyrosine residue at position 341 can be replaced with a phenylalanine residue
  • the methionine at position 347 can be replaced with the glutamic acid, arginine or tryptophane residue
  • the threonine at position 351 can be replaced with the glutamic acid residue
  • the tryptophane residue at position 352 can be replaced with a cysteine, glutamic acid or alanine residue
  • the serine at position 353 can be replaced with the histidine or asparagine residue
  • the leucine at position 354 can be replaced with an alanine residue
  • the lysine at position 360 can be replaced with an alanine or phenylalanine residue of a sequence corresponding to GenBankTM: ACX34092.1.
  • Correspondence of amino acid positions described herein is frequently expressed in terms of the positions of the amino acids of the form of wild
  • said mutant form can comprise at least one mutated amino acid of the wild type calcineurin heterodimer b at positions: V120, N123, L124 or K125, preferably double mutations at positions L124 and K125.
  • the valine at position 120 can be replaced with a serine, an aspartic acid, phenylalanine or leucine residue;
  • the asparagine at position 123 can be replaced with a tryptophan, lysine, phenylalanine, arginine, histidine or serine;
  • the leucine at position 124 can be replaced with a threonine residue;
  • the lysine at position 125 can be replaced with an alanine, a glutamic acid, tryptophan, or two residues such as leucine-arginine or isoleucine-glutamic acid can be added after the lysine at position 125 in the amino acid sequence corresponding to GenBankTM: ACX34095.1.
  • AGT is a DNA repair protein that confers resistance to the cytotoxic effects of alkylating agents, such as nitrosoureas and temozolomide (TMZ).
  • 6-benzylguanine (6-BG) is an inhibitor of AGT that potentiates nitrosourea toxicity and is co-administered with TMZ to potentiate the cytotoxic effects of this agent.
  • 6-BG 6-benzylguanine
  • AGT mutant form can comprise a mutated amino acid of the wild type AGT position P140, in the amino acid sequence such as disclosed in the database Uniprot under the reference P16455.
  • said proline at position 140 is replaced with a lysine residue.
  • MDR1 multidrug resistance protein 1
  • P-GP P-glycoprotein
  • Multidrug Resistance Protein 1 Mediates Resistance to Mitoxantrone via Glutathione-Dependent Drug Efflux Mol Pharmacol April 2006 69:1499-1505).
  • Still another way of preparing drug resistant cells is to prepare cells with specific mutation (s) such as mutations at Arg486 and Glu571 in the Human Topoisomerase II gene, to confer resistance to amsacrine (S. PATEL, B. A. KELLER, and L. M. FISHER. 2000. MOLECULAR PHARMACOLOGY. Vol 57: p 784-791 (2000).
  • specific mutation such as mutations at Arg486 and Glu571 in the Human Topoisomerase II gene
  • Still another way of preparing drug resistant cells is to prepare cells overexpressing microRNA-21 to confer resistance to Daunorubicine (Involvement of miR-21 in resistance to daunorubicin by regulating PTEN expression in the leukaemia K562 cell line Bai, Haitao et al. FEBS Letters, Volume 585, Issue 2, 402-408).
  • cells bearing such a drug resistance conferring mRNA or protein also comprise an inhibitory mRNA or a gene the expression of which is conditioned, allowing the selective destruction of said drug resistant cells in the presence of said drug or upon administration of said drug.
  • Drug resistance gene can also confer resistance to cytotoxic antibiotics, and can be ble gene or mcrA gene. Ectopic expression of ble gene or mcrA in an immune cell gives a selective advantage when exposed to the chemotherapeutic agent, respectively the bleomycine or the mitomycin C.
  • said drug resistance gene can be expressed in the cell by introducing a transgene preferably encoded by at least one vector into a cell.
  • cells bearing a drug resistance gene or a modified gene conferring resistance to a drug also comprise an inducible suicide gene—the induction of which provokes cell death—allowing their selective destruction.
  • the genetic modification step of the method can comprise a step of introduction into cells of an exogeneous nucleic acid comprising at least a sequence encoding the drug resistance gene and a portion of an endogenous gene such that homologous recombination occurs between the endogenous gene and the exogeneous nucleic acid.
  • said endogenous gene can be the wild type “drug resistance” gene, such that after homologous recombination, the wild type gene is replaced by the mutant form of the gene which confers resistance to the drug.
  • the method of the invention further comprises the step of expressing in the cell a rare-cutting endonuclease which is able to cleave a target sequence within an endogenous gene.
  • Said endogenous gene can encode for examples DHFR, IMPDH2, calcineurin or AGT.
  • Said rare-cutting endonuclease can be a TALE-nuclease, a Zinc finger nuclease, a CRISPR/Cas9 endonuclease, a MBBBD-nuclease or a meganuclease.
  • said drug resistance can be conferred to the cell of the invention (anti-CLL1 CAR expressing immune cell,) by the inactivation of a drug sensitizing gene.
  • the inventor sought to inactivate potential drug sensitizing gene to engineer T-cell for immunotherapy, in particular to engineer anti-CLL1 CAR expressing immune cell that can be used in combination with a therapeutic agent (anti-cancer drug).
  • the genetic modification of the method relies on the expression, in provided cells to engineer, of one rare-cutting endonuclease such that said rare-cutting endonuclease specifically catalyzes cleavage in one targeted gene thereby inactivating said targeted gene.
  • the step of inactivating at least one drug sensitizing gene comprises introducing into the cell a rare-cutting endonuclease able to disrupt at least one drug sensitizing gene.
  • said cells are transformed with nucleic acid encoding a rare-cutting endonuclease capable of disrupting a drug sensitizing gene, and said rare-cutting endonuclease is expressed into said cells.
  • Said rare-cutting endonuclease can be a meganuclease, a Zinc finger nuclease, CRISPR/Cas9 nuclease, A MBBBD-nuclease or a TALE-nuclease.
  • said rare-cutting endonuclease is a TALE-nuclease.
  • drug sensitizing gene which can be inactivated to confer drug resistance to the T-cell is the human deoxycytidine kinase (dCK) gene.
  • dCK human deoxycytidine kinase
  • This enzyme is required for the phosphorylation of the deoxyribonucleosides deoxycytidine (dC), deoxyguanosine (dG) and deoxyadenosine (dA).
  • Purine nucleotide analogs PNAs are metabolized by dCK into mono-, di- and tri-phosphate PNA.
  • RNR ribonucleotide reductase
  • the inactivation of dCK in T cells is mediated by TALE nuclease.
  • TALE nuclease a sequence of dCK TALE-nuclease.
  • TALE-nuclease pairs which can be used according to the invention are depicted in PCT/EP2014/075317.
  • PNAs purine nucleoside analogs
  • the dCK inactivation in T cells is combined with an inactivation of TRAC genes rendering these double knock out (KO) T cells both resistant to drug such as clofarabine and less allogeneic.
  • This double features is particularly useful for a therapeutic goal, allowing “off-the-shelf” allogeneic cells for immunotherapy in conjunction with chemotherapy to treat patients with cancer.
  • This double KO inactivation dCK/TRAC can be performed simultaneously or sequentially.
  • TALE-nuclease dCK/TRAC pairs which gave success in the invention is described in PCT/EP2014/075317, in particular, the target sequences in the 2 loci (dCK and TRAC).
  • HPRT human hypoxanthine-guanine phosphoribosyl transferase gene (GenBankTM: M26434.1).
  • HPRT can be inactivated in engineered T-cells to confer resistance to a cytostatic metabolite, the 6-thioguanine (6TG) which is converted by HPRT to cytotoxic thioguanine nucleotide and which is currently used to treat patients with cancer, in particular leukemias (Hacke, Treger et al. 2013).
  • Guanines analogs are metabolized by HPRT transferase that catalyzes addition of phosphoribosyl moiety and enables the formation of TGMP Guanine analogues including 6 mercapthopurine (6MP) and 6 thioguanine (6TG) are usually used as lymphodepleting drugs to treat leukemias. They are metabolized by HPRT (hypoxanthine phosphoribosyl transferase that catalyzes addition of phosphoribosyl moiety and enables formation TGMP. Their subsequent phosphorylations lead to the formation of their triphosphorylated forms that are eventually integrated into DNA. Once incorporated into DNA, thio GTP impairs fidelity of DNA replication via its thiolate groupment and generate random point mutation that are highly deleterious for cell integrity.
  • the present invention provides an anti-CLL1 scCAR expressing cell, in particular an anti-CLL1 scCAR expressing T cell wherein the scCAR has a polypeptide sequence according to SEQ ID NO.35 to 112 (optionally humanized) and wherein the dCK gene is inactivated.
  • the inactivation of the CD3 normally expressed at the surface of the T-cell can confer resistance to anti-CD3 antibodies such as teplizumab.
  • the inventors sought to develop an “off-the shelf” immunotherapy strategy, using allogeneic T-cells, in particular allogenic anti-CLL1 scCAR expressing T-cell resistant to multiple drugs to mediate selection of engineered T-cells when the patient is treated with different drugs.
  • the therapeutic efficiency can be significantly enhanced by genetically engineering multiple drug resistance allogeneic T-cells.
  • Such a strategy can be particularly effective in treating tumors that respond to drug combinations that exhibit synergistic effects.
  • multiple resistant engineered T-cells can expand and be selected using minimal dose of drug agents.
  • the method according to the present invention can comprise modifying T-cell to confer multiple drug resistance to said T-cell.
  • Said multiple drug resistance can be conferred by either expressing more than one drug resistance gene or by inactivating more than one drug sensitizing gene.
  • the multiple drug resistance can be conferred to said T-cell by expressing at least one drug resistance gene and inactivating at least one drug sensitizing gene.
  • the multiple drug resistance can be conferred to said T-cell by expressing at least one drug resistance gene such as mutant form of DHFR, mutant form of IMPDH2, mutant form of calcineurin, mutant form of MGMT, the ble gene, and the mcrA gene and inactivating at least one drug sensitizing gene such as HPRT gene.
  • multiple drug resistance can be conferred by inactivating HPRT gene and expressing a mutant form of DHFR; or by inactivating HPRT gene and expressing a mutant form of IMPDH2; or by inactivating HPRT gene and expressing a mutant form of calcineurin; by inactivating HPRT gene and expressing a mutant form of MGMT; by inactivating HPRT gene and expressing the ble gene; by inactivating HPRT gene and expressing the mcrA gene.
  • the present invention provides allogenic anti-CLL1 scCAR expressing T-cell expressing more than one drug resistance gene or wherein more than one drug sensitizing gene is inactivated.
  • the method of the invention can comprises the transformation of said T-cells with a recombinant suicide gene.
  • Said recombinant suicide gene is used to reduce the risk of direct toxicity and/or uncontrolled proliferation of said T-cells once administrated in a subject (Quintarelli C, Vera F, blood 2007; Tey S K, Dotti G., Rooney C M, boil blood marrow transplant 2007).
  • Suicide genes enable selective deletion of transformed cells in vivo.
  • suicide gene has the ability to convert a non-toxic pro-drug into cytotoxic drug or to express the toxic gene expression product.
  • suicide gene is a nucleic acid coding for a product, wherein the product causes cell death by itself or in the presence of other compounds.
  • a representative example of such a suicide gene is one which codes for thymidine kinase of herpes simplex virus. Additional examples are thymidine kinase of varicella zoster virus and the bacterial gene cytosine deaminase which can convert 5-fluorocytosine to the highly toxic compound 5-fluorouracil.
  • Suicide genes also include as non limiting examples caspase-9 or caspase-8 or cytosine deaminase. Caspase-9 can be activated using a specific chemical inducer of dimerization (CID).
  • Suicide genes can also be polypeptides that are expressed at the surface of the cell and can make the cells sensitive to therapeutic monoclonal antibodies.
  • prodrug means any compound useful in the methods of the present invention that can be converted to a toxic product.
  • the prodrug is converted to a toxic product by the gene product of the suicide gene in the method of the present invention.
  • a representative example of such a prodrug is ganciclovir which is converted in vivo to a toxic compound by HSV-thymidine kinase. The ganciclovir derivative subsequently is toxic to tumor cells.
  • prodrugs include acyclovir, FIAU [1-(2-deoxy-2-fluoro- ⁇ -D-arabinofuranosyl)-5-iodouracil], 6-methoxypurine arabinoside for VZV-TK, and 5-fluorocytosine for cytosine deaminase.
  • One preferred suicide gene system employs a recombinant antigenic polypeptide comprising antigenic motif recognized by the anti-CD20 mAb Rituximab, especially QBen10, such as in the so-called RQR8 polypeptide described in WO2013153391, which is expressed independently from the anti-CLL1 CAR.
  • Rituximab an authorized antibody drug, can then be used for cell depletion when needed.
  • the present invention provides allogenic anti-CLL1 scCAR expressing T-cell expressing more than one drug resistance gene or wherein more than one drug sensitizing gene is inactivated, and a suicide gene allowing said cells to be destroyed.
  • the present invention relates to an allogeneic T-cell, in particular an allogeneic anti-CLL1 scCAR expressing T-cell, and preferably an allogeneic anti-CLL1 scCAR expressing T-cell comprising a peptide having 80% to 100% identity with scfv from SC02-357, SC02-378, SCO2-161, M26, M31, G4, M22, M29, M2, M5, G12, 21.26 or 1075.7 antibodies, said allogeneic anti-CLL1 scCAR expressing T-cell comprising a peptide having 80% to 100% identity with scfv from SC02-357, SC02-378, SCO2-161, M26, M31, G4, M22, M29, M2, M5, G12, 21.26 and 1075.7 antibodies is more particularly resistant to a drug, and specifically suitable for immunotherapy.
  • the resistance of a drug can be conferred by inactivation of drug sensitizing genes or by expression of drug resistance genes.
  • drugs which suit to the invention are the purine nucleoside analogues (PNAs) such as clofarabine or fludarabine, or other drugs such as 6-Mercaptopurine (6MP) and 6 thio-guanine (6TG).
  • the present invention provides methods for engineering immune cells to make them resistant to purine nucleotide analogs (PNA), such a clorofarabine or fludarabine, so that they can be used in cancer immunotherapy treatments in patients pre-treated with these conventional chemotherapies.
  • PNA purine nucleotide analogs
  • the resistance to drugs can be conferred to the T-cells by inactivating one or more gene(s) responsible for the cell's sensitivity to the drug (drug sensitizing gene(s)), such as the dcK and/or HPRT genes.
  • drug sensitizing gene(s) such as the dcK and/or HPRT genes.
  • the resistance to drugs can be conferred to a T-cell by expressing a drug resistance gene.
  • Variant alleles of several genes such as dihydrofolate reductase (DHFR), inosine monophosphate dehydrogenase 2 (IMPDH2), calcineurin or methylguanine transferase (MGMT) have been identified to confer drug resistance to a cell according to the invention.
  • DHFR dihydrofolate reductase
  • IMPDH2 inosine monophosphate dehydrogenase 2
  • MGMT methylguanine transferase
  • CD52 and glucocorticoid receptors which are drug targets of Campath® (alemtuzumab) or rituximab and glucocorticoids treatments, can be inactivated to make the cells resistant to these treatments and give them a competitive advantage over patient's own T-cells not endowed with specific anti-CLL1 scCARs.
  • Expression of CD3 gene can also be suppressed or reduced to confer resistance to Teplizumab, which is another immune suppressive drug.
  • Expression of HPRT can also be suppressed or reduced according to the invention to confer resistance to 6-thioguanine, a cytostatic agent commonly used in chemotherapy especially for the treatment of acute lymphoblasic leukemia.
  • the immune cells can be further manipulated to make them more active or limit exhaustion, by inactivating genes encoding proteins that act as “immune checkpoints” that act as regulators of T-cells activation, such as the following gene selected from CTLA4, PPP2CA, PPP2CB, PTPN6, PTPN22, PDCD1, LAG3, HAVCR2, BTLA, CD160, TIGIT, CD96, CRTAM, LAIR1, SIGLEC7, SIGLEC9, CD244, TNFRSF10B, TNFRSF10A, CASP8, CASP10, CASP3, CASP6, CASP7, FADD, FAS, TGFBRII, TGFBRI, SMAD2, SMAD3, SMAD4, SMAD10, SKI, SKIL, TGIF1, IL 10 RA, IL10RB, HMOX2, IL6R, IL6ST, CSK, PAG1, SIT1, FOXP3, PRDM1 (orbli
  • the present invention also provides allogeneic T-cells expressing an anti-CLL1 scCAR, in particular an anti-CLL1, wherein at least one gene expressing one or more component of T-cell receptor (TCR) is inactivated and/or one gene selected from the genes CTLA4, PPP2CA, PPP2CB, PTPN6, PTPN22, PDCD1, LAG3, HAVCR2, BTLA, CD160, TIGIT, CD96, CRTAM, LAIR1, SIGLEC7, SIGLEC9, CD244, TNFRSF10B, TNFRSF10A, CASP8, CASP10, CASP3, CASP6, CASP7, FADD, FAS, TGFBRII, TGFBRI, SMAD2, SMAD3, SMAD4, SMAD10, SKI, SKIL, TGIF1, IL 10 RA, IL 10 RB, HMOX2, IL6R, IL6ST, CSK, PAG1, SIT1, FOXP3, PRDM
  • said gene is a gene that acts as a regulator of T-cells activation coding the beta 2 microglobulin protein.
  • the anti-CLL1 scCAR-immune cells of the invention can be further manipulated to make them resistant to a drug, in particular to a drug used during chemotherapy against cancer, in particular a CLL1-expressing cell-mediated cancer such as AML.
  • a drug in particular to a drug used during chemotherapy against cancer, in particular a CLL1-expressing cell-mediated cancer such as AML.
  • This can be achieved by introducing a gene conferring resistance to said drug.
  • This same gene may be turned on and off by using a gene inducible inhibition/expression system as previously described (Garcia E L, Mills A A (2002) Getting around lethality with inducible Cre-mediated excision. Semin Cell Dev Biol 13:151-8, Lewandoski M (2001) Conditional control of gene expression in the mouse.
  • anti-CLL1 scCAR-expressing, drug resistant immune cell wherein (i) at least one gene expressing one or more component of T-cell receptor (TCR) is inactivated (ii) at least one gene conferring resistance to a drug is incorporated or a gene conferring sensitivity to said drug is deleted or mutated to be inactivated (iii) optionally another gene selected from the gene disclosed in the following table 9 is inactivated—is an object of the present invention.
  • TCR T-cell receptor
  • the present invention encompasses the isolated anti-CLL1 scCAR-immune cells or cell lines obtainable by the method of the invention, more particularly isolated cells comprising any of the proteins, polypeptides, allelic variants, altered or deleted genes or vectors described herein.
  • the immune cells of the present invention or cell lines can further comprise exogenous recombinant polynucleotides, in particular scCARs or suicide genes or they can comprise altered or deleted genes coding for checkpoint proteins or ligands thereof that contribute to their efficiency as a therapeutic product, ideally as an “off the shelf” product.
  • the present invention concerns the method for treating or preventing cancer in the patient by administrating at least once an engineered immune cell obtainable by the above methods.
  • the present invention encompasses also a method for manufacturing target cells which express both a surface receptor specific to the scCAR T cells and a resistance gene. These target cells are particularly useful for testing the cytotoxicity of scCAR T cells. These cells are readily resistant to clinically relevant dose of clofarabine and harbor luciferase activity. This combination of features enable traking them in vivo in a mice model or destroy them when required.
  • Clofarabine resistant Daudi cells mimick the physiological state of acute myeloma leukemia (AML) patients relapsing form induction therapy, that harbor drug resistant B cell malignancies. Thus, these cells are of great interest to evaluate the reliability and cytotoxicity of drug resistant scCAR T cells.
  • these target cells are CLL1+ Luciferase+ Daudi cells.
  • An anti-CLL1 CAR of the invention may include at least the insertion of at least one epitope in the extracellular domain of said CAR. This is intended to temptatively deplete the immune cells endowed with the CAR in the event of in vivo adverse effects such as a cytokine storm. Moreover, such insertion of epitope or “epitope-tagging” may be useful to sort or purify the engineered immune cells in-vitro during their manufacturing process Said at least one epitope may be any antigenic peptide which is enough immunogenic to be bound by a specific antibody recognizing such peptide.
  • this can be obtained, for instance, by inserting at least one, and preferably two copies of a CD20 mimotope, preferably of sequence CPYSNPSLCS (SEQ ID NO.113), into the CAR polypeptide sequence.
  • a CD20 mimotope preferably of sequence CPYSNPSLCS (SEQ ID NO.113)
  • SEQ ID NO.113 sequence CPYSNPSLCS
  • the order of the scFvs from the N terminal end to the C terminal end is presented as follows: the VH chain and then the VL chain.
  • this order is inversed: VL chain and then the VL chain.
  • Different positions of the at least one CD20 mimotope within the anti-CLL1 CAR of the invention are schematized in FIG. 3 .
  • Said two copies of a CD20 mimotope can be linked to each other and also to the V L by a linker. They can also be inserted between the anti-CLL1 scFv and the hinge (such as CD8alpha), by using an optional linker.
  • the CD20 mimotopes can be bound by anti-CD20 antibodies, such as Rituximab (McLaughlin P, et al. 1998).
  • the anti-CLL1 CAR of the present invention may comprise VH and a VL chains which are able to bind to CLL1 cell surface antigen, optionally humanized, a linker L, a suicide domain, a hinge or part of it, a transmembrane domain, a co-stimulatory domain and a stimulatory domain.
  • the epitope is a mimotope.
  • the mimotope As a macromolecule, often a peptide, which mimics the structure of an epitope, the mimotope has the advantage to be smaller than conventional epitope, and therefore may be beneficial for a non-conformational sequence and easier to reproduce in a long polypeptide such a CAR.
  • Mimotopes are known for several pharmaceutically-approved mAb such as two 10 amino acid peptides for cetuximab (Riemer et al., 2005), or a 24 aa for palivizumab (Arbiza et al, 1992). As these mimotopes can be identified by phage display, it is possible to try several of them in order to obtain a sequence which does not perturb the scFv for the same mAb. Furthermore, their use can enhance a complement-dependentcytotoxicity (CDC).
  • CDC complement-dependentcytotoxicity
  • the epitope introduced within the chimeric scFv is the CD20 mimotope (SEQ ID NO.113) and the infused mAb presenting an affinity to this mimotope—for sorting and/or depletion purpose(s)—is rituximab.
  • said at least one epitope is inserted between the VH and VL chains of the anti-CLL1.1 CAR, optionally linked to said VH and VL chains by one linker.
  • the term “linker” as used in the context of a scFv refers to a peptide linker that consists of amino acids such as glycine and/or serine residues used alone or in combination, to link variable heavy and variable light chain regions together.
  • the flexible polypeptide linker is a Glycine/Serine linker and comprises the amino acid sequence (Gly-Gly-Gly-Ser) n or (Gly-Gly-Gly-Gly-Ser) n , where n is a positive integer equal to or greater than 1.
  • the flexible polypeptide linkers include, but are not limited to, (Gly 4 Ser) 4 or (Gly 4 Ser) 3 .
  • said CLL1 specific chimeric antigen receptor has one of the polypeptide structure selected from V1, V3 or V5, as illustrated in FIG. 2 , wherein one CD20 mimotope is inserted between the VH and VL chains of the anti-CLL1.1 CAR, optionally linked to said VH and VL chains by one linker.
  • said CLL1 specific chimeric antigen receptor has one of the polypeptide structure of version V3 as illustrated in FIG. 2 , said structure comprising at least an anti-CLL1 extra cellular ligand binding-domain, Cd8 ⁇ transmembrane domain, 4-1BB co-stimulatory domain, CD3 zeta signaling domain, and wherein one CD20 mimotope is inserted between the VH and VL chains of the anti-CLL1.1 CAR, optionally linked to said VH and VL chains by one linker.
  • said at least one epitope is inserted at the N terminal end of the CAR—so upfront of the scFvs—, optionally linked to the VH chain and to the N terminal end of the CAR by one linker.
  • said CLL1 specific chimeric antigen receptor has one of the polypeptide structure selected from V1, V3 or V5, as illustrated in FIG. 2 , said structure comprising at least an anti-CLL1 extra cellular ligand binding-domain, Cd8 ⁇ transmembrane domain, 4-1BB co-stimulatory domain, CD3 zeta signaling domain, and wherein one epitope is inserted at the N terminal end of the CAR—so upfront of the scFvs—, optionally linked to the VH chain and to the N terminal end of the CAR by one linker.
  • said CLL1 specific chimeric antigen receptor has one of the polypeptide structure of version V3 as illustrated in FIG. 2 , said structure comprising at least an anti-CLL1 extra cellular ligand binding-domain, Cd8 ⁇ transmembrane domain, 4-1BB co-stimulatory domain, CD3 zeta signaling domain, and wherein one epitope is inserted at the N terminal end of the CAR—so upfront of the scFvs—, optionally linked to the VH chain and to the N terminal end of the CAR by one linker.
  • said at least one epitope is inserted between the scFvs and the hinge of the CAR, optionally linked to the VL chain and to the hinge by one linker.
  • said CLL1 specific chimeric antigen receptor has one of the polypeptide structure selected from V1, V3 or V5, as illustrated in FIG. 2 , said structure comprising at least an anti-CLL1 extra cellular ligand binding-domain, Cd8 ⁇ transmembrane domain, 4-1BB co-stimulatory domain, CD3 zeta signaling domain, and wherein one epitope is inserted between the scFvs and the hinge of the CAR, optionally linked to the VL chain and to the hinge by one linker.
  • said CLL1 specific chimeric antigen receptor has one of the polypeptide structure of version V3 as illustrated in FIG. 2 , said structure comprising at least an anti-CLL1 extra cellular ligand binding-domain, Cd8 ⁇ transmembrane domain, 4-1BB co-stimulatory domain, CD3 zeta signaling domain, and wherein one epitope is inserted between the scFvs and the hinge of the CAR, optionally linked to the VL chain and to the hinge by one linker.
  • At least two epitopes are inserted in the extracellular domain of the anti-CLL1 CAR of the present invention.
  • CLL1 specific chimeric antigen receptor has one of the polypeptide structure selected from V1, V3 or V5, as illustrated in FIG. 2 , said structure comprising at least an anti-CLL1 extra cellular ligand binding-domain, Cd8 ⁇ transmembrane domain, 4-1BB co-stimulatory domain, CD3 zeta signaling domain, and two CD20 mimotopes,
  • said extra-binding domain comprising VH and VL chains directed against CLL1 and a Fc ⁇ RIII ⁇ or Cd8 ⁇ or IgG1 hinge;
  • linker SEQ ID NO.10 being interspaced between the 2 epitopes and/or between the VH and the 2 epitopes.
  • CLL1 specific chimeric antigen receptor has one of the polypeptide structure selected from V1, V3 or V5, as illustrated in FIG. 2 , said structure comprising at least an anti-CLL1 extra cellular ligand binding-domain, Cd8 ⁇ transmembrane domain, 4-1BB co-stimulatory domain, CD3 zeta signaling domain, and two CD20 mimotopes,
  • said extra-binding domain comprising VH and VL chains directed against CLL1 and a Fc ⁇ RIII ⁇ or Cd8 ⁇ or IgG1 hinge;
  • linker SEQ ID NO.10
  • SEQ ID NO.10 being interspaced between the 2 epitopes and/or at the N terminal end of the CAR.
  • At least two epitopes are inserted in the extracellular domain in such a way that the VH is located between them, all these components being optionally interspaced by at least one linker.
  • said CLL1 specific chimeric antigen receptor has one of the polypeptide structure selected from V1, V3 or V5, as illustrated in FIG. 2 , said structure comprising at least an anti-CLL1 extra cellular ligand binding-domain, Cd8 ⁇ transmembrane domain, 4-1BB co-stimulatory domain, CD3 zeta signaling domain, and wherein two epitopes are inserted in the extracellular domain in such a way that the VH is located between them, all these components being optionally interspaced by at least one linker.
  • said CLL1 specific chimeric antigen receptor has one of the polypeptide structure of version V3 as illustrated in FIG. 2 , said structure comprising at least an anti-CLL1 extra cellular ligand binding-domain, Cd8 ⁇ transmembrane domain, 4-1BB co-stimulatory domain, CD3 zeta signaling domain, and wherein two epitopes are inserted in the extracellular domain in such a way that the VL is located between them, all these components being optionally interspaced by at least one linker.
  • two epitopes are inserted in the extracellular domain in such a way that the VL is located between them, all these components being optionally interspaced by at least one linker.
  • said CLL1 specific chimeric antigen receptor has one of the polypeptide structure selected from V1, V3 or V5, as illustrated in FIG. 2 , said structure comprising at least an anti-CLL1 extra cellular ligand binding-domain, Cd8 ⁇ transmembrane domain, 4-1BB co-stimulatory domain, CD3 zeta signaling domain, and wherein two epitopes are inserted in the extracellular domain in such a way that the VL is located between them, all these components being optionally interspaced by at least one linker.
  • said CLL1 specific chimeric antigen receptor has one of the polypeptide structure of version V3 as illustrated in FIG. 2 , said structure comprising at least an anti-CLL1 extra cellular ligand binding-domain, Cd8 ⁇ transmembrane domain, 4-1BB co-stimulatory domain, CD3 zeta signaling domain, and wherein two epitopes are inserted in the extracellular domain in such a way that the VL is located between them, all these components being optionally interspaced by at least one linker.
  • said CLL1 specific chimeric antigen receptor comprises an extracellular binding domain wherein at least two epitopes are inserted in the extracellular domain in such a way that the VH and VL chains ar located between them, all these components being optionally interspaced by at least one linker.
  • said CLL1 specific chimeric antigen receptor has one of the polypeptide structure selected from V1, V3 or V5, as illustrated in FIG. 2 , said structure comprising at least an anti-CLL1 extra cellular ligand binding-domain, Cd8 ⁇ transmembrane domain, 4-1BB co-stimulatory domain, CD3 zeta signaling domain, and wherein two epitopes are inserted in the extracellular domain in such a way that the VH and VL chains ar located between them, all these components being optionally interspaced by at least one linker.
  • said CLL1 specific chimeric antigen receptor has one of the polypeptide structure of version V3 as illustrated in FIG. 2 , said structure comprising at least an anti-CLL1 extra cellular ligand binding-domain, Cd8 ⁇ transmembrane domain, 4-1BB co-stimulatory domain, CD3 zeta signaling domain, and wherein two epitopes are inserted in the extracellular domain in such a way that the VH and VL chains ar located between them, all these components being optionally interspaced by at least one linker.
  • three epitopes are inserted in the extracellular domain of the anti-CLL1 CAR of the present invention.
  • said CLL1 specific CAR of the invention contains an extracellular binding domain wherein three epitopes are inserted in the extracellular domain in such a way that the VH and VL chains ar located between them, all these components being optionally interspaced by at least one linker.
  • said CLL1 specific chimeric antigen receptor has one of the polypeptide structure selected from V1, V3 or V5, as illustrated in FIG. 2 , said structure comprising at least an anti-CLL1 extra cellular ligand binding-domain, Cd8 ⁇ transmembrane domain, 4-1BB co-stimulatory domain, CD3 zeta signaling domain, and wherein three epitopes are inserted in the extracellular domain in such a way that the VH and VL chains ar located between them, all these components being optionally interspaced by at least one linker.
  • said CLL1 specific chimeric antigen receptor has one of the polypeptide structure of version V3 as illustrated in FIG. 2 , said structure comprising at least an extracellular ligand binding-domain anti-CLL1, Cd8 ⁇ transmembrane domain, 4-1BB co-stimulatory domain, CD3 zeta signaling domain, and wherein three epitopes are inserted in the extracellular domain in such a way that the VH and VL chains ar located between them, all these components being optionally interspaced by at least one linker.
  • CLL1 specific chimeric antigen receptor has one of the polypeptide structure selected from V1, V3 or V5, as illustrated in FIG. 2 , said structure comprising at least an anti-CLL1 extra cellular ligand binding-domain, Cd8 ⁇ transmembrane domain, 4-1BB co-stimulatory domain, CD3 zeta signaling domain, and three CD20 epitopes,
  • said extra-binding domain comprising VH and VL chains directed against CLL1 and a Fc ⁇ RIII ⁇ or Cd8 ⁇ or IgG1 hinge;
  • linker SEQ ID NO.10 being interspaced between the 3 epitopes and/or between the VH and the 3 epitopes.
  • CLL1 specific chimeric antigen receptor has one of the polypeptide structure selected from V1, V3 or V5, as illustrated in FIG. 2 , said structure comprising at least an anti-CLL1 extra cellular ligand binding-domain, Cd8 ⁇ transmembrane domain, 4-1BB co-stimulatory domain, CD3 zeta signaling domain, two CD20 epitopes, and one CD34 epitope;
  • said extra-binding domain comprising VH and VL chains directed against CLL1 and a Fc ⁇ RIII ⁇ or Cd8 ⁇ or IgG1 hinge;
  • CD34 epitope being inserted between the said 2 CD20 epitopes, all components being interspaced between them by a linker (SEQ ID NO.10) and a linker between the epitope and and between the VH and the 3 epitopes.
  • the VH and VL chains which are used as extracellular binding domain are binding preferably to human membrane CLL1-1.
  • said above anti-CLL1 CARs comprising at least an extra cellular ligand binding-domain including VH and VL chains derived from anti-CLL1 monoclonal antibodies.
  • epitopes can be included into the CAR of the present invention such as follows:
  • the extracellular binding domain comprises at least 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 mAb-specific epitopes.
  • the extracellular binding domain comprises at least 1, 2 or 3 mAb-specific epitopes.
  • the extracellular binding domain comprises several mAb-specific epitopes
  • all the mAb-specific epitopes are identical.
  • the extracellular binding domain when the extracellular binding domain comprises several mAb-specific epitopes, the mAb-specific epitopes are not identical.
  • the extracellular binding domain can comprises three mAb-specific epitopes, two of them being identical and the third one being different.
  • the extracellular binding domain comprises a VH, a VL, one or more mAb-specific epitopes, preferably 1, 2 or 3, more preferably 2 or 3 mAb-specific epitopes.
  • the extracellular binding domain comprises the following sequence (Nterm is located on the left hand side):
  • V 1 and V 2 are V H and V L of an ScFv (i.e., V 1 is V L and V 2 is V H or V 1 is V H and V 2 is V L );
  • L 1 is any linker suitable to link the VH chain to the VL chain in an ScFv;
  • L is a linker, preferably comprising glycine and serine residues, and each occurrence of L in the extracellular binding domain can be identical or different to other occurrence of L in the same extracellular binding domain, and,
  • x is 0 or 1 and each occurrence of x is independently from the others;
  • epitope 1 epitope 2 and epitope 3 are mAb-specific epitopes and can be identical or different.
  • the extracellular binding domain comprises the following sequence (Nterm is located on the left hand side):
  • L 1 is a linker comprising Glycine and/or Serine. In some embodiment, L 1 is a linker comprising the amino acid sequence (Gly-Gly-Gly-Ser) n or (Gly-Gly-Gly-Gly-Ser) n , where n is 1, 2, 3, 4 or 5. In some embodiments L 1 is (Gly 4 Ser) 4 or (Gly4Ser) 3 .
  • L is a flexible linker, preferably comprising Glycine and/or Serine.
  • L has an amino acid sequence selected from SGG, GGS, SGGS (SEQ ID NO. 200), SSGGS (SEQ ID NO. 201), GGGG (SEQ ID NO. 202), SGGGG (SEQ ID NO. 203), GGGGS (SEQ ID NO. 204), SGGGGS (SEQ ID NO. 205), GGGGGS (SEQ ID NO. 206), SGGGGGS (SEQ ID NO. 207), SGGGGG (SEQ ID NO. 208), GSGGGGS (SEQ ID NO. 209), GGGGGGGS (SEQ ID NO.
  • SGGGGGGG SEQ ID NO. 211
  • SGGGGGGGS SEQ ID NO. 212
  • SGGGGSGGGGS SEQ ID NO. 213 preferably SGG, SGGS (SEQ ID NO. 200), SSGGS (SEQ ID NO. 201), GGGG (SEQ ID NO. 202), SGGGGS (SEQ ID NO. 205), SGGGGGS (SEQ ID NO. 207), SGGGGG (SEQ ID NO. 208), GSGGGGS (SEQ ID NO. 209), or SGGGGSGGGGS (SEQ ID NO. 213).
  • all the Ls are identical.
  • the extracellular binding domain comprises several occurrences of L
  • the Ls are not all identical.
  • L is SGGGGS (SEQ ID NO. 205).
  • the extracellular binding domain comprises several occurrences of L and all the Ls are SGGGGS (SEQ ID NO. 205).
  • Epitope 1, Epitope 2 and Epitope 3 are identical or different and are selected from mAb-specific epitopes having an amino acid sequence of anyone of SEQ ID NO 33 to SEQ ID NO 42.
  • Epitope 1, Epitope 2 and Epitope 3 are identical or different and are selected from mAb-specific epitopes specifically recognized by ibritumomab, tiuxetan, muromonab-CD3, tositumomab, abciximab, basiliximab, brentuximab vedotin, cetuximab, infliximab, rituximab, alemtuzumab, bevacizumab, certolizumab pegol, daclizumab, eculizumab, efalizumab, gemtuzumab, natalizumab, omalizumab, palivizumab, ranibizumab, tocilizumab, trastuzumab, vedolizumab, adalimumab, belimumab, canakinumab, denosumab, golimumab, i
  • Epitope 1 is an mAb-specific epitope having an amino acid sequence of SEQ ID NO 33.
  • Epitope 2 is an mAb-specific epitope having an amino acid sequence of SEQ ID NO 35.
  • Epitope 3 is an mAb-specific epitope having an amino acid sequence of SEQ ID NO 36.
  • Epitope 4 is an mAb-specific epitope having an amino acid sequence of SEQ ID NO 37.
  • Epitope 4 is an mAb-specific epitope having an amino acid sequence of SEQ ID NO 38.
  • Epitope 2 is an mAb-specific epitope having an amino acid sequence of SEQ ID NO 33 and Epitope 3 is an mAb-specific epitope having an amino acid sequence of SEQ ID NO 41 or 42.
  • one of Epitope 1, Epitope 2, Epitope 3 and Epitope 4 is a CD34 epitope, preferably an epitope of SEQ ID 41 or 42.
  • one of Epitope1, Epitope 2, Epitope 3 and Epitope 4 is a CD34 epitope, preferably an epitope of SEQ ID 41 or 42 and the other mAb specific epitopes are CD20 mimotopes, preferably mimotope of SEQ ID NO 33.
  • the immune cells expressing the CLL1 sepcific CAR according to the present invention may comprise epitope(s) in their extracellular domain such as described above, so that they can be depleted in a patient in the event of adverse or too acute immune response (e.g. cytokine storm) by administering to said patient an antibody-preferably monoclonal-specific to said epitope (s).
  • adverse or too acute immune response e.g. cytokine storm
  • in vivo depletion is meant in the present invention the administration of a treatment to a mammalian organism aiming to stop the proliferation of CAR-expressing immune cells by inhibition or elimination.
  • One aspect of the invention is related to a method for in vivo depleting an engineered immune cell expressing a CAR comprising an m-Ab specific epitope as previously described, comprising contacting said engineered immune cell or said CAR-expressing immune cell with at least one epitope-specific mAbs.
  • Another aspect of the invention relates to a method for in vivo depleting immune CAR-expressing immune cell which comprises the above chimeric scFv (formed by insertion of a mAb-specific epitope) by contacting said engineered immune cell with epitope-specific antibodies.
  • said immune cells are T-cells and/or the antibodies are monoclonal.
  • the in vivo depletion of immune engineered cell is performed on engineered immune cell which has been previously sorted using the in vitro method of the present invention. In this case, this will be the same infused mAb used.
  • the mAb-specific antigen is CD20 antigen and the epitope-specific mAb is rituximab.
  • the invention relates to a method for in vivo depleting an engineered immune cell expressing a CAR comprising an mAb-specific epitope (CAR-expressing immune cell) as previously described, in a patient comprising contacting said CAR-expressing immune cell with at least one epitope-specific mAbs.
  • CAR-expressing immune cell mAb-specific epitope
  • said mAb-specific epitope is a CD20 epitope or mimotope, preferably SEQ ID NO 35 and the epitope-specific mAbs is rituximab.
  • the step of contacting said engineered immune cell or said CAR-expressing immune cell with at least one epitope-specific mAb comprises infusing the patient with epitope-specific mAb, preferably rituximab.
  • the amount of viable CAR-expressing immune cells decreases, preferably by at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80% or 90%.
  • the CDC assay is the assay disclosed in Example 3, Example 4 or Example 7.4.
  • said mAb-specific epitope is a CD20 epitope or mimotope, preferably SEQ ID NO 35 and the epitope-specific mAbs is rituximab.
  • the epitopes inserted into the extracellular domain of the CARs may be useful to the steps of sorting or purifying the immune cells expressing said CARs, as part of the method for producing them.
  • the resulting cells are engineered immune cell expressing at the cell surface membrane a CLL1 specific chimeric antigen receptor as previously described, in particular engineered immune cells derived from primary T-lymphocytes, optionally resistant to an anti-cancer drug, and bearing a deletion in a gene coding for an alpha TCR or a beta TCR.
  • the present invention discloses an engineered immune cell as above, wherein expression of TCR is suppressed.
  • the present invention discloses an engineered immune cell as above, wherein expression of at least one MHC protein, preferably ⁇ 2m or HLA, is reduced or suppressed in said engineered immune cell.
  • ⁇ 2m stands for beta 2 microglobulin and HLA for human leukocyte antigen.
  • the MHC protein is a MHC protein of Class I or of class II.
  • the present invention discloses an engineered immune cell as above, wherein said engineered immune cell is mutated to confer resistance to at least one immune suppressive drug, chemotherapy drug, or anti-cancer drug.
  • the present invention discloses an engineered immune cell as above for use in therapy.
  • the present invention discloses an engineered immune cell for use in therapy as above, wherein the patient is a human.
  • the present invention discloses an engineered immune cell for use in therapy as above, wherein the condition is a pre-malignant or malignant cancer condition characterized by CLL1-expressing cells.
  • the present invention discloses an engineered immune cell for use in therapy as above, wherein the condition is a condition which is characterized by an overabundance of CLL1-expressing cells.
  • the present invention discloses an engineered immune cell for use in therapy as above, wherein the malignant cancer condition is a hematological cancer condition.
  • the present invention discloses an engineered immune cell for use in therapy as above, wherein the hematological cancer condition is leukemia or malignant lymphoproliferative disorders.
  • the present invention discloses an engineered immune cell for use in therapy as above, wherein said leukemia is selected from the group consisting of acute myelogenous leukemia, chronic myelogenous leukemia, myelodysplastic syndrome, acute lymphoid leukemia, chronic lymphoid leukemia, and myelodysplastic syndrome.
  • the present invention discloses an engineered immune cell for use in therapy as above, wherein the leukemia is acute myelogenous leukemia (AML).
  • AML acute myelogenous leukemia
  • the present invention discloses an engineered immune cell for use in therapy as above, wherein said hematologic cancer is a malignant lymphoproliferative disorder.
  • the present invention discloses an engineered immune cell for use in therapy as above, wherein said malignant lymphoproliferative disorder is lymphoma.
  • the present invention discloses an engineered immune cell for use in therapy as above, wherein said lymphoma is selected from the group consisting of multiple myeloma, non-Hodgkin's lymphoma, Burkitt's lymphoma, and follicular lymphoma (small cell and large cell).
  • the present invention discloses a method of impairing a hematologic cancer cell comprising contacting said hematologic cancer cell with an engineered cell, which at least expresses anti-CLL1 CAR such as exposed above, in an amount effective to cause impairment of said cancer cell.
  • the present invention thus discloses a method of engineering an immune cell comprising:
  • the present invention discloses a method of engineering an immune cell as above comprising:
  • the present invention discloses a method of engineering an immune cell as above comprising:
  • the present invention discloses a method of treating a subject in need thereof comprising:
  • the present invention discloses a method of treating a subject in need thereof as above, wherein said immune cell is provided from a donor.
  • the present invention discloses a method of treating a subject in need thereof as above, wherein said immune cell is provided from the patient himself.
  • isolated cell obtained by the different methods or cell line derived from said isolated cell as previously described can be used as a medicament.
  • said medicament can be used for treating cancer, particularly for the treatment of leukemia in a patient in need thereof.
  • said isolated cell according to the invention or cell line derived from said isolated cell can be used in the manufacture of a medicament for treatment of a cancer in a patient in need thereof.
  • an anti-CLL1 CAR expressing T cell is provided as a medicament for the treatment of AML, of an AML subtype, of an AML-related complication, of an AML-related condition.
  • said medicament can be used for treating a CLL1-expressing cell-mediated pathological condition or a condition characterized by the direct or indirect activity of a CLL1-expressing cell.
  • the present invention relies on methods for treating patients in need thereof, said method comprising at least one of the following steps:
  • said T cells of the invention can undergo robust in vivo T cell expansion and can persist for an extended amount of time.
  • Said treatment can be ameliorating, curative or prophylactic. It may be either part of an autologous immunotherapy or part of an allogenic immunotherapy treatment.
  • autologous it is meant that cells, cell line or population of cells used for treating patients are originating from said patient or from a Human Leucocyte Antigen (HLA) compatible donor.
  • HLA Human Leucocyte Antigen
  • allogeneic is meant that the cells or population of cells used for treating patients are not originating from said patient but from a donor.
  • Said treatment can be used to treat patients diagnosed wherein a pre-malignant or malignant cancer condition characterized by CLL1-expressing cells, especially by an overabundance of CLL1-expressing cells. Such conditions are found in hematologic cancers, such as leukemia.
  • the present invention provides a composition for its use in the treatment of a CLL1 expressing cells-mediated disease, in particular a CLL1 expressing cells—mediated hematologic cancer, said composition comprising said anti-CLL1 scCAR expressing T cell of the invention.
  • Any other CLL1-mediating or CLL1-involving malignant lymphoproliferative disorders disclosed herein may be improved with the anti-CLL1 CAR-expressing cells of the present invention.
  • the cancer that may be treated using the anti-CLL1 CAR-expressing cells of the present invention is leukemia, a disease associated to leukemia or a complication thereof.
  • Leukemias that can be treated using the anti-CLL1 CAR-expressing cells of the present invention can be acute myelogenous leukemia (AML).
  • AML or AML subtypes that may be treated using the anti-CLL1 scCAR-expressing cells of the present invention may be in particular, acute myeloblastic leukemia, minimally differentiated acute myeloblastic leukemia, acute myeloblastic leukemia without maturation, acute myeloblastic leukemia with granulocytic maturation, promyelocytic or acute promyelocytic leukemia (APL), acute myelomonocytic leukemia, myelomonocytic together with bone marrow eosinophilia, acute monoblastic leukemia (M5a) or acute monocytic leukemia (M5b), acute erythroid leukemias, including erythroleukemia (M6a) and very rare pure erythroid leukemia (M6
  • Subtypes of AML also include, hairy cell leukemia, philadelphia chromosome-positive acute lymphoblastic leukemia.
  • AML may be classified as AML with specific genetic abnormalities. Classification is based on the ability of karyotype to predict response to induction therapy, relapse risk, survival.
  • AML that may be treated using the anti-CLL1 CAR-expressing cells of the present invention may be AML with a translocation between chromosomes 8 and 21, AML with a translocation or inversion in chromosome 16, AML with a translocation between chromosomes 9 and 11, APL (M3) with a translocation between chromosomes 15 and 17, AML with a translocation between chromosomes 6 and 9, AML with a translocation or inversion in chromosome 3, AML (megakaryoblastic) with a translocation between chromosomes 1 and 22.
  • the present invention is particularly useful for the treatment of AML associated with these particular cytogenetic markers.
  • the present invention also provides an anti-CLL1 CAR expressing T cell for the treatment of patients with specific cytogenetic subsets of AML, such as patients with t(15;17)(q22;q21) identified using all-trans retinoic acid (ATRA)16-19 and for the treatment of patients with t(8;21)(q22;q22) or inv(16)(p13q22)/t(16;16)(p13;q22) identified using repetitive doses of high-dose cytarabine.
  • ATRA all-trans retinoic acid
  • the present invention provides an anti-CLL1 CAR expressing T cell for the treatment of patients with aberrations, such as ⁇ 5/del(5q), ⁇ 7, abnormalities of 3q, or a complex karyotype, who have been shown to have inferior complete remission rates and survival.
  • aberrations such as ⁇ 5/del(5q), ⁇ 7, abnormalities of 3q, or a complex karyotype, who have been shown to have inferior complete remission rates and survival.
  • chemotherapeutic agent refers to a medicament, preferably a compound or a derivative thereof that can interact with a cancer cell, thereby reducing the proliferative status of the cell and/or killing the cell.
  • chemotherapeutic agents or “anti-cancer drug” include, but are not limited to, alkylating agents (e.g., busulfan, carboplatine, chlorambucil, cisplatine, cyclophosphamide, ifosfamide, melphalan, mechlorethamine, oxaliplatine, uramustine, temozolomide, fotemustine), metabolic antagonists (e.g., purine nucleoside antimetabolite such as clofarabine, fludarabine or T-deoxyadenosine, methotrexate (MTX), 5-fluorouracil or derivatives thereof, azathioprine, capecitabine, cytarabine, floxuridine, fluorouracile, gemcitabine, methotrexate, pemetrexed), antitumor antibiotics (e.g., mitomycin, adriamycin, bleomycine, daunorubicine, doxor
  • a therapeutic agent refers to a compound or a derivative thereof that may be used to treat cancer, in particular to treat a hematopoietic cancer cell and more particularly AML, thereby reducing the proliferative status of the cancer cell and/or killing the cancer cell.
  • chemotherapeutic agents include, but are not limited to aracytine, Cytosine arabinoside, amsacrine, daunorubicine, idarubicine, novantrone, mitoxantrone, vepeside, etoposide (VP16), arsenic trioxyde, transretinoic acid, mechlorethamine, procarbazine, chlorambucil, and combination thereof.
  • cells of the invention are administered to a patient in conjunction with a drug (or an agent) selected from aracytine, cytosine arabinoside, amsacrine, daunorubicine, idarubicine, novantrone, mitoxantrone, vepeside, etoposide (VP16), arsenic trioxyde, transretinoic acid, cytarabine, anthracyclines, 6-thioguanine, hydroxyurea, prednisone, and combination thereof.
  • a drug or an agent selected from aracytine, cytosine arabinoside, amsacrine, daunorubicine, idarubicine, novantrone, mitoxantrone, vepeside, etoposide (VP16), arsenic trioxyde, transretinoic acid, cytarabine, anthracyclines, 6-thioguanine, hydroxyurea, prednisone, and combination thereof.
  • Such agents may further include, but are not limited to, the anti-cancer agents TRIMETHOTRIXATETM (TMTX), TEMOZOLOMIDETM, RALTRITREXEDTM, S-(4-Nitrobenzyl)-6-thioinosine (NBMPR), 6-benzyguanidine (6-BG), bis-chloronitrosourea (BCNU) and CAMPTOTHECINTM, or a therapeutic derivative of any thereof.
  • TTTX TRIMETHOTRIXATETM
  • TEMOZOLOMIDETM TEMOZOLOMIDETM
  • RALTRITREXEDTM S-(4-Nitrobenzyl)-6-thioinosine
  • NBMPR 6-benzyguanidine
  • BCNU bis-chloronitrosourea
  • CAMPTOTHECINTM CAMPTOTHECINTM
  • an anti-CLL1 scCAR expressing T cell is administered to a patient, in combination with at least one therapeutic agent selected from aracytine, Cytosine arabinoside, amsacrine, daunorubicine, idarubicine, novantrone, mitoxantrone, vepeside, etoposide (VP16), arsenic trioxyde, transretinoic acid and combination thereof.
  • at least one therapeutic agent selected from aracytine, Cytosine arabinoside, amsacrine, daunorubicine, idarubicine, novantrone, mitoxantrone, vepeside, etoposide (VP16), arsenic trioxyde, transretinoic acid and combination thereof.
  • a cell which is “resistant or tolerant” to an agent means a cell which has been genetically modified so that the cell proliferates in the presence of an amount of an agent that inhibits or prevents proliferation of a cell without the modification.
  • the invention provides a treatment for AML in patients over 60 years or in patients of less than 20 years.
  • the present invention provides a pediatric treatment, in particular a pediatric treatment against AML, or AML-related diseases or complications.
  • the present invention is used as a treatment in AML patients with low, poor or unfavorable status that is to say with a predicted survival of less than 5 years survival rate.
  • patients suffering AML with the following cytogenetic characteristics: ⁇ 5; 5q; ⁇ 7; 7q-;11q23; non t(9;11); inv(3); t(3;3); t(6;9); t(9;22) is associated with poor-risk status (Byrd J. C. et al., Dec. 15, 2002; Blood: 100 (13) and is especially contemplated to be treated according to the present invention or with an object of the present invention.
  • the anti-CLL1 CAR expressing T cell of present invention may be used as induction therapy, as post remission therapy of AML or as a consolidation therapy in patient with AML.
  • the anti-CLL1 CAR expressing T cell of the present invention may be used in case of AML relapse, or in case of refractory or resistant AML, and more preferably, in combination with at least one other anti-cancer drug
  • At least one anti-CLL1 CAR expressing cell of the invention is used for preventing cancer cells development occurring in particular after anti-cancer treatment, during bone marrow depletion or before bone marrow transplantation, after bone marrow destruction.
  • the invention provides a medicament that improves the health condition of a patient, in particular a patient undergoing a complication related to AML. More preferably, said engineered anti-CLL1 CAR expressing T cell of the invention is expressing at least one anti-CLL1 CAR of the invention and is used as a medicament for the treatment of a complication related to AML.
  • a complication or disease related to AML may include a preceding myelodysplasia phase, secondary leukemia, in particular secondary AML, high white blood cell count, and absence of Auer rods.
  • secondary leukemia in particular secondary AML
  • high white blood cell count and absence of Auer rods.
  • CNS central nervous system
  • hyperleukocytosis residual disease
  • the present invention also provides an anti-CLL1 CAR expressing T cell for the treatment of a pathological condition related to AML.
  • the present invention provides a therapy for AML related myeloid neoplasms, for acute myeloid leukemia and myelodysplastic syndrome, a treatment of relapsed or refractory acute myeloid leukemia, a treatment of relapsed or refractory acute promyelocytic leukemia in adults, a treatment for acute promyeloid leukaemia, a treatment of acute myeloid leukemia in adults over 60 years.
  • the present invention provides a composition for the treatment of AML associated diseases, in particular hematologic malignancy related to AML.
  • MDS myelodysplasia syndromes
  • preleukemia myelodysplasia syndromes
  • genetic syndromes associated with the risk of AML can be improved with the adequate use of the present invention
  • said genetic syndromes include Down syndrome, trisomy, Fanconi anemia, Bloom syndrome, Ataxia-telangiectasia, Diamond-Blackfan anemia, Schwachman-Diamond syndrome, Li-Fraumeni syndrome, Neurofibromatosis type 1, Severe congenital neutropenia (also called Kostmann syndrome).
  • the present invention provides a composition for the treatment of CLL1+cell-mediated diseases.
  • CLL1+cell mediated diseases include inflammation, such as rheumatoid arthitis.
  • the present invention can be used for the treatment of CLL1+cell mediated diseases such as inflammation and more particularly rheumatoid arthritis.
  • the present invention also provides a composition comprising an engineered T cells according to the invention for its use or a method for treating a disease.
  • the disease is a hematologic cancer, in particular a stem cell cancer including but is not limited to leukemia (such as acute myelogenous leukemia (AML) or a complication thereof.
  • leukemia such as acute myelogenous leukemia (AML) or a complication thereof.
  • the present invention also provides a composition for its use or a method for inhibiting the proliferation or reducing a CLL1-expressing cell population or activity in a patient.
  • An exemplary method includes contacting a population of cells comprising a CLL1-expressing cell with an anti-CLL1 CART cell, and in particular scCART, of the invention that binds to the CLL1-expressing cell.
  • the present invention provides a composition for its use or a method for inhibiting the proliferation or reducing the population of cancer cells expressing CLL1 in a patient, the methods comprising contacting the CLL1-expressing cancer cell population with an anti-CLL1 CART cell, and in particular scCART, of the invention that binds to the CLL1-expressing cell, binding of an anti-CLL1 CAR cell, and in particular scCART, of the invention to the CLL1-expressing cancer cell resulting in the destruction of the CLL1-expressing cancer cells
  • the anti-CLL1 CART cell, and in particular scCART, of the invention reduces the quantity, number, amount or percentage of cells and/or cancer cells by at least 25%, at least 30%, at least 40%, at least 50%, at least 65%, at least 75%, at least 85%, at least 95%, or at least 99% (to undetectable level) in a subject with or animal model for myeloid leukemia or another cancer associated with CLL1-expressing cells, relative to a negative control.
  • the present invention also provides a composition for its use or a method for preventing, treating and/or managing a disorder or condition associated with CLL1-expressing cells (e.g., associated with a hematologic cancer), the methods comprising administering to a subject in need an anti-CLL1 CART cell, and in particular scCART, of the invention that binds to the CLL1-expressing cell.
  • the subject is a human.
  • disorders associated with CLL1-expressing cells include inflammatory disorders (such as rheumatoid arthritis) and cancers (such as hematological cancers, in particular AML or AML complications).
  • the present invention also provides a composition for its use or a method for preventing, treating and/or managing a disease associated with CLL1-expressing cells, the method comprising administering to a subject in need an anti-CLL1 CART cell, and in particular scCART, of the invention that binds to the CLL1-expressing cell.
  • the subject is a human.
  • diseases associated with CLL1-expressing cells include in particular Acute Myeloid Leukemia (AML).
  • the present invention provides a composition for its use or a method for treating or preventing relapse of cancer associated with CLL1-expressing cells, the method comprising administering to a subject in need thereof an anti-CLL1 CART cell, and in particular scCART, of the invention that binds to the CLL1-expressing cell.
  • the methods comprise administering to the subject in need thereof an effective amount of an anti CLL1 CART cell, and in particular scCART, of the invention that binds to the CLL1-expressing cell in combination with an effective amount of another therapy.
  • CLL1 is considered to be a “cancer stem cell” marker in AML. Therefore, an anti-CLL1 CART cell, and in particular scCART, of the invention can prevent relapse of AML, or even treat AML that is mostly CLL1-negative but with a “stem” population of CLL1+ cells (a CLL1-expressing cells).
  • the invention provides compositions and methods for treating subjects that have undergone treatment for a disease or disorder associated with elevated expression levels of CD 19, and exhibits a disease or disorder associated with elevated levels of CLL1.
  • the treatment with the engineered immune cells according to the invention may be in combination with one or more therapies against cancer selected from the group of antibodies therapy, chemotherapy, cytokines therapy, dendritic cell therapy, gene therapy, hormone therapy, laser light therapy and radiation therapy.
  • the treatment with the engineered immune cells according to the invention may be administered in combination (e.g., before, simultaneously or following) with one or more therapies against cancer selected from aracytine, cytosine arabinoside, amsacrine, daunorubicine, idarubicine, novantrone, mitoxantrone, vepeside, etoposide (VP16), arsenic trioxyde, transretinoic acid, combination of arsenic trioxyde, transretinoic acid, mechlorethamine, procarbazine, chlorambucil, and combination thereof.
  • one or more therapies against cancer selected from aracytine, cytosine arabinoside, amsacrine, daunorubicine, idarubicine, novantrone, mitoxantrone, vepeside, etoposide (VP16), arsenic trioxyde, transretinoic acid, combination of arsenic trioxyde, transretinoic acid, mechlorethamine, proc
  • said treatment can be administrated into patients undergoing an immunosuppressive treatment.
  • the present invention preferably relies on cells or population of cells, which have been made resistant to at least one immunosuppressive agent due to the inactivation of a gene encoding a receptor for such immunosuppressive agent.
  • the immunosuppressive treatment should help the selection and expansion of the T-cells according to the invention within the patient.
  • the administration of the cells or population of cells according to the present invention may be carried out in any convenient manner, including by aerosol inhalation, injection, ingestion, transfusion, implantation or transplantation.
  • the compositions described herein may be administered to a patient subcutaneously, intradermaly, intratumorally, intranodally, intramedullary, intramuscularly, by intravenous or intralymphatic injection, or intraperitoneally.
  • the cell compositions of the present invention are preferably administered by intravenous injection.
  • the administration of the cells or population of cells can consist of the administration of 10 4 -10 9 cells per kg body weight, preferably 10 5 to 10 6 cells/kg body weight including all integer values of cell numbers within those ranges.
  • the cells or population of cells can be administrated in one or more doses.
  • said effective amount of cells are administrated as a single dose.
  • said effective amount of cells are administrated as more than one dose over a period time. Timing of administration is within the judgment of managing physician and depends on the clinical condition of the patient.
  • the cells or population of cells may be obtained from any source, such as a blood bank or a donor. While individual needs vary, determination of optimal ranges of effective amounts of a given cell type for a particular disease or conditions within the skill of the art.
  • An effective amount means an amount which provides a therapeutic or prophylactic benefit.
  • the dosage administrated will be dependent upon the age, health and weight of the recipient, kind of concurrent treatment, if any, frequency of treatment and the nature of the effect desired.
  • said effective amount of cells or composition comprising those cells are administrated parenterally.
  • Said administration can be an intravenous administration.
  • Said administration can be directly done by injection within a tumor.
  • cells are administered to a patient in conjunction with (e.g., before, simultaneously or following) any number of relevant treatment modalities, including but not limited to treatment with agents such as antiviral therapy, cidofovir and interleukin-2, Cytarabine (also known as ARA-C) or natalizimab treatment for MS patients or efaliztimab treatment for psoriasis patients or other treatments for PML patients.
  • agents such as antiviral therapy, cidofovir and interleukin-2, Cytarabine (also known as ARA-C) or natalizimab treatment for MS patients or efaliztimab treatment for psoriasis patients or other treatments for PML patients.
  • the T cells of the invention may be used in combination with chemotherapy, radiation, immunosuppressive agents, such as cyclosporin, azathioprine, methotrexate, mycophenolate, and FK506, antibodies, or other immunoablative agents such as CAMPATH, anti-CD3 antibodies or other antibody therapies, cytoxin, fludaribine, cyclosporin, FK506, rapamycin, mycoplienolic acid, steroids, FR901228, cytokines, and irradiation.
  • immunosuppressive agents such as cyclosporin, azathioprine, methotrexate, mycophenolate, and FK506, antibodies
  • immunoablative agents such as CAMPATH, anti-CD3 antibodies or other antibody therapies
  • cytoxin fludaribine
  • cyclosporin FK506, rapamycin
  • mycoplienolic acid steroids
  • steroids FR901228
  • cytokines irradiation
  • the cell compositions of the present invention are administered to a patient in conjunction with (e.g., before, simultaneously or following) bone marrow transplantation, T cell ablative therapy using either chemotherapy agents such as, fludarabine, external-beam radiation therapy (XRT), cyclophosphamide, or antibodies such as OKT3 or CAMPATH.
  • chemotherapy agents such as, fludarabine, external-beam radiation therapy (XRT), cyclophosphamide, or antibodies such as OKT3 or CAMPATH.
  • the cell compositions of the present invention are administered following B-cell ablative therapy such as agents that react with CD20, e.g., Rituxan.
  • B-cell ablative therapy such as agents that react with CD20, e.g., Rituxan.
  • subjects may undergo standard treatment with high dose chemotherapy followed by peripheral blood stem cell transplantation.
  • subjects receive an infusion of the expanded immune cells of the present invention.
  • expanded cells are administered before or following surgery.
  • anti-CLL1 scCAR expressing cells are administered to a patient in conjunction (e.g., before, simultaneously or following) with a drug selected from aracytine, cytosine arabinoside, amsacrine, daunorubicine, idarubicine, novantrone, mitoxantrone, vepeside, etoposide (VP16), arsenic trioxyde, transretinoic acid, combination of arsenic trioxyde, transretinoic acid, mechlorethamine, procarbazine, chlorambucil, and combination thereof.
  • a drug selected from aracytine, cytosine arabinoside, amsacrine, daunorubicine, idarubicine, novantrone, mitoxantrone, vepeside, etoposide (VP16), arsenic trioxyde, transretinoic acid, combination of arsenic trioxyde, transretinoic acid, mechlorethamine, procarbazin
  • anti-CLL1 scCAR expressing cells are administered to a patient in conjunction with a drug selected from cytarabine, anthracyclines, 6-thioguanine, hydroxyurea, prednisone, and combination thereof.
  • a,” “an,” “the,” and “at least one” are used interchangeably and mean one or more than one.
  • Amino acid residues in a polypeptide sequence are designated herein according to the one-letter code, in which, for example, Q means Gln or Glutamine residue, R means Arg or Arginine residue and D means Asp or Aspartic acid residue.
  • Amino acid substitution means the replacement of one amino acid residue with another, for instance the replacement of an Arginine residue with a Glutamine residue in a peptide sequence is an amino acid substitution.
  • Nucleotides are designated as follows: one-letter code is used for designating the base of a nucleoside: A is adenine, T is thymine, C is cytosine, and G is guanine.
  • r represents g or a (purine nucleotides)
  • k represents g or t
  • s represents g or c
  • w represents a or t
  • m represents a or c
  • y represents t or c (pyrimidine nucleotides)
  • d represents g, a or t
  • v represents g, a or c
  • b represents g, t or c
  • h represents a, t or c
  • n represents g, a, t or c.
  • nucleic acid or “polynucleotides” refers to nucleotides and/or polynucleotides, such as deoxyribonucleic acid (DNA) or ribonucleic acid (RNA), oligonucleotides, fragments generated by the polymerase chain reaction (PCR), and fragments generated by any of ligation, scission, endonuclease action, and exonuclease action.
  • DNA deoxyribonucleic acid
  • RNA ribonucleic acid
  • PCR polymerase chain reaction
  • Nucleic acid molecules can be composed of monomers that are naturally-occurring nucleotides (such as DNA and RNA), or analogs of naturally-occurring nucleotides (e.g., enantiomeric forms of naturally-occurring nucleotides), or a combination of both.
  • Modified nucleotides can have alterations in sugar moieties and/or in pyrimidine or purine base moieties.
  • Sugar modifications include, for example, replacement of one or more hydroxyl groups with halogens, alkyl groups, amines, and azido groups, or sugars can be functionalized as ethers or esters.
  • sugar moiety can be replaced with sterically and electronically similar structures, such as aza-sugars and carbocyclic sugar analogs.
  • modifications in a base moiety include alkylated purines and pyrimidines, acylated purines or pyrimidines, or other well-known heterocyclic substitutes.
  • Nucleic acid monomers can be linked by phosphodiester bonds or analogs of such linkages. Nucleic acids can be either single stranded or double stranded.
  • delivery vector or “delivery vectors” is intended any delivery vector which can be used in the present invention to put into cell contact (i.e. “contacting”) or deliver inside cells or subcellular compartments (i.e. “introducing”) agents/chemicals and molecules (proteins or nucleic acids) needed in the present invention. It includes, but is not limited to liposomal delivery vectors, viral delivery vectors, drug delivery vectors, chemical carriers, polymeric carriers, lipoplexes, polyplexes, dendrimers, microbubbles (ultrasound contrast agents), nanoparticles, emulsions or other appropriate transfer vectors.
  • delivery vectors allow delivery of molecules, chemicals, macromolecules (genes, proteins), or other vectors such as plasmids, peptides developed by Diatos. In these cases, delivery vectors are molecule carriers.
  • delivery vector or “delivery vectors” is also intended delivery methods to perform transfection.
  • vector refers to a nucleic acid molecule capable of transporting another nucleic acid to which it has been linked.
  • a “vector” in the present invention includes, but is not limited to, a viral vector, a plasmid, a RNA vector or a linear or circular DNA or RNA molecule which may consists of a chromosomal, non chromosomal, semi-synthetic or synthetic nucleic acids.
  • Preferred vectors are those capable of autonomous replication (episomal vector) and/or expression of nucleic acids to which they are linked (expression vectors). Large numbers of suitable vectors are known to those of skill in the art and commercially available.
  • Viral vectors include retrovirus, adenovirus, parvovirus (e. g. adenoassociated viruses), coronavirus, negative strand RNA viruses such as orthomyxovirus (e. g., influenza virus), rhabdovirus (e. g., rabies and vesicular stomatitis virus), paramyxovirus (e. g. measles and Sendai), positive strand RNA viruses such as picornavirus and alphavirus, and double-stranded DNA viruses including adenovirus, herpesvirus (e. g., Herpes Simplex virus types 1 and 2, Epstein-Barr virus, cytomegalovirus), and poxvirus (e.
  • orthomyxovirus e. g., influenza virus
  • rhabdovirus e. g., rabies and vesicular stomatitis virus
  • paramyxovirus e. g. measles and Sendai
  • viruses include Norwalk virus, togavirus, flavivirus, reoviruses, papovavirus, hepadnavirus, and hepatitis virus, for example.
  • retroviruses include: avian leukosis-sarcoma, mammalian C-type, B-type viruses, D type viruses, HTLV-BLV group, lentivirus, spumavirus (Coffin, J. M., Retroviridae: The viruses and their replication, In Fundamental Virology, Third Edition, B. N. Fields, et al., Eds., Lippincott-Raven Publishers, Philadelphia, 1996).
  • lentiviral vector HIV-Based lentiviral vectors that are very promising for gene delivery because of their relatively large packaging capacity, reduced immunogenicity and their ability to stably transduce with high efficiency a large range of different cell types.
  • Lentiviral vectors are usually generated following transient transfection of three (packaging, envelope and transfer) or more plasmids into producer cells.
  • lentiviral vectors enter the target cell through the interaction of viral surface glycoproteins with receptors on the cell surface.
  • the viral RNA undergoes reverse transcription, which is mediated by the viral reverse transcriptase complex.
  • the product of reverse transcription is a double-stranded linear viral DNA, which is the substrate for viral integration in the DNA of infected cells.
  • integrated lentiviral vectors or LV
  • NILV non-integrative lentiviral vectors
  • Delivery vectors and vectors can be associated or combined with any cellular permeabilization techniques such as sonoporation or electroporation or derivatives of these techniques.
  • cell or cells any eukaryotic living cells, primary cells and cell lines derived from these organisms for in vitro cultures.
  • primary cell or “primary cells” are intended cells taken directly from living tissue (i.e. biopsy material) and established for growth in vitro, that have undergone very few population doublings and are therefore more representative of the main functional components and characteristics of tissues from which they are derived from, in comparison to continuous tumorigenic or artificially immortalized cell lines.
  • cell lines can be selected from the group consisting of CHO-K1 cells; HEK293 cells; Caco2 cells; U2-OS cells; NIH 3T3 cells; NSO cells; SP2 cells; CHO—S cells; DG44 cells; K-562 cells, U-937 cells; MRC5 cells; IMR90 cells; Jurkat cells; HepG2 cells; HeLa cells; HT-1080 cells; HCT-116 cells; Hu-h7 cells; Huvec cells; Molt 4 cells.
  • All these cell lines can be modified by the method of the present invention to provide cell line models to produce, express, quantify, detect, study a gene or a protein of interest; these models can also be used to screen biologically active molecules of interest in research and production and various fields such as chemical, biofuels, therapeutics and agronomy as non-limiting examples.
  • mutant is intended the substitution, deletion, insertion of up to one, two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, fifteen, twenty, twenty five, thirty, fourty, fifty, or more nucleotides/amino acids in a polynucleotide (cDNA, gene) or a polypeptide sequence.
  • the mutation can affect the coding sequence of a gene or its regulatory sequence. It may also affect the structure of the genomic sequence or the structure/stability of the encoded mRNA.
  • variant(s) it is intended a repeat variant, a variant, a DNA binding variant, a TALE-nuclease variant, a polypeptide variant obtained by mutation or replacement of at least one residue in the amino acid sequence of the parent molecule.
  • “functional variant” is intended a catalytically active mutant of a protein or a protein domain; such mutant may have the same activity compared to its parent protein or protein domain or additional properties, or higher or lower activity.
  • identity refers to sequence identity between two nucleic acid molecules or polypeptides. Identity can be determined by comparing a position in each sequence which may be aligned for purposes of comparison. When a position in the compared sequence is occupied by the same base, then the molecules are identical at that position. A degree of similarity or identity between nucleic acid or amino acid sequences is a function of the number of identical or matching nucleotides at positions shared by the nucleic acid sequences. Various alignment algorithms and/or programs may be used to calculate the identity between two sequences, including FASTA, or BLAST which are available as a part of the GCG sequence analysis package (University of Wisconsin, Madison, Wis.), and can be used with, e.g., default setting.
  • polypeptides having at least 70%, 85%, 90%, 95%, 98% or 99% identity to specific polypeptides described herein and preferably exhibiting substantially the same functions, as well as polynucleotide encoding such polypeptides, are contemplated.
  • similarity describes the relationship between the amino acid sequences of two or more polypeptides.
  • BLASTP may also be used to identify an amino acid sequence having at least 70%, 75%, 80%, 85%, 87.5%, 90%, 92.5%, 95%, 97.5%, 98%, 99% sequence similarity to a reference amino acid sequence using a similarity matrix such as BLOSUM45, BLOSUM62 or BLOSUM80. Unless otherwise indicated a similarity score will be based on use of BLOSUM62.
  • BLOSUM45 BLOSUM45
  • BLOSUM62 BLOSUM80
  • BLASTP “Identities” shows the number and fraction of total residues in the high scoring sequence pairs which are identical; and BLASTP “Positives” shows the number and fraction of residues for which the alignment scores have positive values and which are similar to each other.
  • Amino acid sequences having these degrees of identity or similarity or any intermediate degree of identity of similarity to the amino acid sequences disclosed herein are contemplated and encompassed by this disclosure.
  • the polynucleotide sequences of similar polypeptides are deduced using the genetic code and may be obtained by conventional means.
  • a functional variant of pTalpha can have 70%, 75%, 80%, 85%, 87.5%, 90%, 92.5%, 95%, 97.5%, 98%, 99% sequence similarity to the amino acid sequence of SEQ ID NO: 107 such as disclosed in WO2013176916.
  • a polynucleotide encoding such a functional variant would be produced by reverse translating its amino acid sequence using the genetic code.
  • subject or “patient” as used herein includes all members of the animal kingdom including non-human primates and humans.
  • relapsed refers to a situation where a subject or a mammal, who has had a remission of cancer after therapy has a return of cancer cells.
  • refractory or resistant refers to a circumstance where a subject or a mammal, even after intensive treatment, has residual cancer cells in his body.
  • drug resistance refers to the condition when a disease does not respond to the treatment of a drug or drugs.
  • Drug resistance can be either intrinsic (or primary resistance), which means the disease has never been responsive to the drug or drugs, or it can be acquired, which means the disease ceases responding to a drug or drugs that the disease had previously responded to (secondary resistance).
  • drug resistance is intrinsic.
  • the drug resistance is acquired.
  • hematologic malignancy refers to a cancer of the body's blood-bone marrow and/or lymphatic tissue.
  • hematological malignancies include, in particular, acute myeloid leukemia (AML), AML with trilineage myelodysplasia (AML/TMDS), mixed lineage leukemia (MLL), and other AM-related pathologies.
  • leukemia refers to malignant neoplasms of the blood-forming tissues, including, in particular to acute myeloid leukemia or acute myelogenous leukemia (AML).
  • AML acute myeloid leukemia
  • T cells were purified from Buffy coat samples provided by EFS (Etableau Francais du Sang, Paris, France) using Ficoll gradient density medium. The PBMC layer was recovered and T cells were purified using a commercially available T-cell enrichment kit. Purified T cells were activated in X-VivoTM-15 medium (LonzaLONZA®) supplemented with 20 ng/mL Human IL-2, 5% Human, and Dynabeads Human T activator CD3/CD28 at a bead:cell ratio 1:1 (Life Technologies).
  • Transfections were done at Day 4 or Day 11 after T-cell purification and activation. 5 millions of cells were transfected with 15 ⁇ g of mRNA encoding the different scCAR constructs.
  • scCAR mRNAs were produced using T7 mRNA polymerase transfections done using Cytopulse technology, by applying two 0.1 mS pulses at 3000V/cm followed by four 0.2 mS pulses at 325V/cm in 0.4 cm gap cuvettes in a final volume of 200 ⁇ l of “Cytoporation buffer T” (BTX Harvard Apparatus). Cells were immediately diluted in X-VivoTM-15 media and incubated at 37° C. with 5% CO 2 . IL-2 was added 2 h after electroporation at 20 ng/mL.
  • T-cells were incubated in 96-well plates (40,000 cells/well), together with an equal amount of cells expressing various levels of the CLL1 protein.
  • Co-cultures were maintained in a final volume of 100 ⁇ l of X-VivoTM-15 medium (LONZA®) for 6 hours at 37° C. with 5% CO 2 .
  • CD107a staining was done 20 during cell stimulation, by the addition of a fluorescent anti-CD107a antibody at the beginning of the co-culture, together with 1 ⁇ g/ml of anti-CD49d, 1 ⁇ g/ml of anti-CD28, and 1 ⁇ Monensin solution.
  • T-cells were incubated in 96-well plates (40,000 cells/well), together with cell lines expressing various levels of the CLL1 protein. Co-cultures were maintained in a final volume of 100 ⁇ l of X-VivoTM-15 medium (LONZA®) for 24 hours at 37° C. with 5% CO 2 . After this incubation period the plates were centrifuged at 1500 rpm for 5 minutes and the supernatants were recovered in a new plate. IFN gamma detection in the cell culture supernatants was done by ELISA assay. The IFN gamma release assays were carried by starting the cell co-cultures 24 h after mRNA transfection.
  • X-VivoTM-15 medium LONZA®
  • T-cells were incubated in 96-well plates (100,000 cells/well), together with 10,000 target cells (expressing CLL1) and 10,000 control (CLL1neg) cells in the same well.
  • Target and control cells were labelled with fluorescent intracellular dyes (CFSE or Cell Trace Violet) before co-culturing them with scCAR+ T-cells.
  • the co-cultures were incubated for 4 hours at 37° C. with 5% CO 2 . After this incubation period, cells were labelled with a fixable viability dye and analyzed by flow cytometry. Viability of each cellular population (target cells or CLL1neg control cells) was determined and the % of specific cell lysis was calculated. Cytotoxicity assays were carried out 48 h after mRNA transfection.
  • scCAR detection at the surface of T-cells was done using a recombinant protein consisting on the fusion of the extracellular domain of the human CLL1 protein, together with a murine IgG1 Fc fragment. Binding of this protein to the scCAR molecule was detected with a fluorochrome-conjugated secondary antibody targeting the mouse Fc portion of the protein, and analyzed by flow cytometry.
  • Immunodeficient NOG mice were intravenously (iv) injected with (CLL1 expressing_MOLM13-Luciferase cells as an AML xenograft mouse model.
  • mice received an anti-cancer treatment.
  • Mice were then iv injected (either 2 or 7 days after injection of the tumor cell line) with different doses of scCAR+ T-cells to be tested, or with T-cells that were not transduced with the scCAR lentiviral vector.
  • Bioluminescent signals were determined at the day of T-cell injection (DO), at D7, 14, 21, 28 and 40 after T-cell injection in order to follow tumoral progression on the different animals.
  • TALE-nuclease targeting two 17-bp long sequences were designed and produced. Each half target is recognized by repeats of the half TALE-nucleases listed in Table 10.
  • TALE-nuclease construct was subcloned using restriction enzyme digestion in a mammalian expression vector under the control of the T7 promoter.
  • mRNA encoding TALE-nuclease cleaving TRAC genomic sequence were synthesized from plasmid carrying the coding sequence downstream from the T7 promoter.
  • T cells preactivated during 72 hours with antiCD3/CD28 coated beads were transfected with each of the 2 mRNAs encoding both half TRAC_T01 TALE-nucleases.
  • different groups of T cells from the same donor were respectively transduced with a lentiviral vector encoding one of the anti-CLL1 scCAR previously described (SEQ ID NO: 35 to 112).
  • CD3 NEG cells were purified using anti-CD3 magnetic beads and 5 days post-transduction cells were reactivated with soluble anti-CD28 (5 ⁇ g/ml).
  • the expression of the activation marker CD25 are analyzed by FACS 7 days post transduction.
  • the purified cells transduced with the lentiviral vector encoding CLL1 scCAR assayed for CD25 expression at their surface in order to assess their activation in comparison with the non-transduced cells.
  • Increased CD25 expression is expected both in CD28 reactivation or no reactivation conditions.
  • T cells were purified from Buffy coat samples provided by EFS (Etableau Francais du Sang, Paris, France) using Ficoll gradient density medium (Ficoll Paque PLUS/GE Healthcare Life Sciences). The PBMC layer was recovered and T cells were purified using a commercially available T-cell enrichment kit (Stem Cell Technologies). Purified T cells were activated in X-VivoTM-15 medium (Lanza) supplemented with 20 ng/mL Human IL-2 (Miltenyi Biotech), 5% Human Serum (Sera Laboratories), and Dynabeads Human T activator CD3/CD28 at a bead:cell ratio 1:1 (Life Technologies). After activation cells were grown and maintained in X-VivoTM-15 medium (LONZA®) supplemented with 20 ng/mL Human IL-2 (Miltenyi Biotec) and 5% Human Serum (Sera Laboratories),
  • Transfections were done at Day 4 or Day 11 after T-cell purification and activation. 5 millions of cells were transfected with 15 ⁇ g of mRNA encoding the different scCAR constructs.
  • scCAR mRNAs were produced using the mMESSAGE mMACHINE® T7 Kit (Life Technologies) and purified using RNeasy® Mini Spin Columns (QIAGEN®). Transfections were done using Cytopulse technology, by applying two 0.1 mS pulses at 3000V/cm followed by four 0.2 mS pulses at 325V/cm in 0.4 cm gap 20 cuvettes in a final volume of 200 ⁇ l of “Cytoporation buffer T” (BTX Harvard Apparatus).
  • IL-2 from Miltenyi Biotec was added 2 h after electroporation at 20 ng/ml.
  • T-cells were incubated in 96-well plates (40,000 cells/well), together with an equal amount of cells expressing or not the CLL1 protein.
  • Co-cultures were maintained in a final volume of 100 ⁇ l of X-VivoTM-15 medium (LONZA®) for 6 hours at 37° C. with 5% CO 2 .
  • CD107a staining was done during cell stimulation, by the addition of a fluorescent anti-CD107a antibody (APC conjugated, from Miltenyi Biotec) at the beginning of the co-culture, together with 1 ⁇ g/ml of anti-CD49d (BD Pharmingen), 1 ⁇ g/ml of anti-CD28 (Miltenyi Biotec), and 1 ⁇ Monensin solution (eBioscience).
  • T-cells were incubated in 96-well plates (40,000 cells/well), together with cell lines expressing or not the CLL1 protein. Co-cultures were maintained in a final volume of 100 ⁇ l of X-VivoTM-15 medium (LONZA®) for 24 hours at 37° C. with 5% CO 2 . After this incubation period the plates were centrifuged at 1500 rpm for 5 minutes and the supernatants were recovered in a new plate. IFN gamma detection in the cell culture supernatants was done by ELISA assay (Human IFN-gamma QUANTIKINE® ELISA Kit, from R&D Systems). The IFN gamma release assays were carried by starting the cell co-cultures 24 h after mRNA transfection.
  • T-cells were incubated in 96-well plates (100,000 cells/well), together with 10,000 target cells (expressing CLL1) and 10,000 control (CLL1neg) cells in the same well.
  • Target and control cells were labelled with fluorescent intracellular dyes (CFSE or Cell Trace Violet, from Life Technologies) before co-culturing them with scCAR+ T-cells.
  • the co-cultures were incubated for 4 hours at 37° C. with 5% CO 2 . After this incubation period, cells were labelled with a fixable viability dye (eFluor 780, from eBioscience) and analyzed by flow cytometry. Viability of each cellular population (target cells or CLL1neg control cells) was determined and the % of specific cell lysis was calculated. Cytotoxicity assays were carried out 48 h after mRNA transfection.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Immunology (AREA)
  • Organic Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Engineering & Computer Science (AREA)
  • Biochemistry (AREA)
  • Zoology (AREA)
  • Biomedical Technology (AREA)
  • Medicinal Chemistry (AREA)
  • Cell Biology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Molecular Biology (AREA)
  • Biotechnology (AREA)
  • Wood Science & Technology (AREA)
  • Biophysics (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Public Health (AREA)
  • Animal Behavior & Ethology (AREA)
  • Veterinary Medicine (AREA)
  • Epidemiology (AREA)
  • Microbiology (AREA)
  • Hematology (AREA)
  • General Engineering & Computer Science (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Toxicology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Oncology (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • General Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Physics & Mathematics (AREA)
  • Mycology (AREA)
  • Analytical Chemistry (AREA)
  • General Physics & Mathematics (AREA)
  • Dispersion Chemistry (AREA)
  • Pathology (AREA)
  • Developmental Biology & Embryology (AREA)
US15/546,633 2015-01-26 2016-01-25 Anti-CLL1 specific single-chain chimeric antigen receptors (scCARs) for cancer immunotherapy Active 2036-07-04 US11014989B2 (en)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
DKPA201570044 2015-01-26
DKPA201570044 2015-01-26
PCT/EP2016/051469 WO2016120218A1 (en) 2015-01-26 2016-01-25 ANTI-CLL1 SPECIFIC SINGLE-CHAIN CHIMERIC ANTIGEN RECEPTORS (scCARS) FOR CANCER IMMUNOTHERAPY

Publications (2)

Publication Number Publication Date
US20180009895A1 US20180009895A1 (en) 2018-01-11
US11014989B2 true US11014989B2 (en) 2021-05-25

Family

ID=52396337

Family Applications (6)

Application Number Title Priority Date Filing Date
US15/546,633 Active 2036-07-04 US11014989B2 (en) 2015-01-26 2016-01-25 Anti-CLL1 specific single-chain chimeric antigen receptors (scCARs) for cancer immunotherapy
US15/546,604 Abandoned US20180000914A1 (en) 2015-01-26 2016-01-25 Anti-hsp70 specific chimeric antigen receptors (cars) for cancer immunotherapy
US15/546,619 Abandoned US20180002427A1 (en) 2015-01-26 2016-01-25 Cll1-specific multi-chain chimeric antigen receptor
US15/546,623 Abandoned US20180051089A1 (en) 2015-01-26 2016-01-25 T cell receptor knock out engineered immune cells, endowed with chimeric antigen receptors binding to cd123 for the treatment of relapsed/refractory acute myeloid lymphoma or blastic plasmacytoid dendritic cell neoplasm
US15/546,630 Abandoned US20180002435A1 (en) 2015-01-26 2016-01-25 mAb-DRIVEN CHIMERIC ANTIGEN RECEPTOR SYSTEMS FOR SORTING/DEPLETING ENGINEERED IMMUNE CELLS
US18/481,797 Pending US20240228636A1 (en) 2015-01-26 2023-10-05 mAb-DRIVEN CHIMERIC ANTIGEN RECEPTOR SYSTEMS FOR SORTING/DEPLETING ENGINEERED IMMUNE CELLS

Family Applications After (5)

Application Number Title Priority Date Filing Date
US15/546,604 Abandoned US20180000914A1 (en) 2015-01-26 2016-01-25 Anti-hsp70 specific chimeric antigen receptors (cars) for cancer immunotherapy
US15/546,619 Abandoned US20180002427A1 (en) 2015-01-26 2016-01-25 Cll1-specific multi-chain chimeric antigen receptor
US15/546,623 Abandoned US20180051089A1 (en) 2015-01-26 2016-01-25 T cell receptor knock out engineered immune cells, endowed with chimeric antigen receptors binding to cd123 for the treatment of relapsed/refractory acute myeloid lymphoma or blastic plasmacytoid dendritic cell neoplasm
US15/546,630 Abandoned US20180002435A1 (en) 2015-01-26 2016-01-25 mAb-DRIVEN CHIMERIC ANTIGEN RECEPTOR SYSTEMS FOR SORTING/DEPLETING ENGINEERED IMMUNE CELLS
US18/481,797 Pending US20240228636A1 (en) 2015-01-26 2023-10-05 mAb-DRIVEN CHIMERIC ANTIGEN RECEPTOR SYSTEMS FOR SORTING/DEPLETING ENGINEERED IMMUNE CELLS

Country Status (14)

Country Link
US (6) US11014989B2 (enExample)
EP (5) EP3250607A1 (enExample)
JP (5) JP2018504143A (enExample)
KR (3) KR102661969B1 (enExample)
CN (5) CN107406516B (enExample)
AU (5) AU2016212158B2 (enExample)
BR (2) BR112017015453A2 (enExample)
CA (5) CA2973525C (enExample)
DK (2) DK3250604T3 (enExample)
ES (2) ES2842212T3 (enExample)
IL (3) IL252937B (enExample)
MX (3) MX2017009349A (enExample)
RU (3) RU2017129455A (enExample)
WO (5) WO2016120216A1 (enExample)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US12054531B2 (en) 2020-03-20 2024-08-06 Lyell Immunopharma, Inc. Recombinant cell surface markers
US12144827B2 (en) 2021-02-25 2024-11-19 Lyell Immunopharma, Inc. ROR1 targeting chimeric antigen receptor

Families Citing this family (163)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US9458450B2 (en) 2012-03-15 2016-10-04 Flodesign Sonics, Inc. Acoustophoretic separation technology using multi-dimensional standing waves
US10967298B2 (en) 2012-03-15 2021-04-06 Flodesign Sonics, Inc. Driver and control for variable impedence load
US10704021B2 (en) 2012-03-15 2020-07-07 Flodesign Sonics, Inc. Acoustic perfusion devices
US9950282B2 (en) 2012-03-15 2018-04-24 Flodesign Sonics, Inc. Electronic configuration and control for acoustic standing wave generation
CA2935960C (en) 2014-01-08 2023-01-10 Bart Lipkens Acoustophoresis device with dual acoustophoretic chamber
PT3119807T (pt) 2014-03-19 2019-09-04 Cellectis Recetores de antigénios quiméricos específicos de cd123 para imunoterapêutica do cancro
SG11201707089WA (en) 2015-04-13 2017-10-30 Pfizer Chimeric antigen receptors targeting b-cell maturation antigen
US11377651B2 (en) 2016-10-19 2022-07-05 Flodesign Sonics, Inc. Cell therapy processes utilizing acoustophoresis
US11708572B2 (en) 2015-04-29 2023-07-25 Flodesign Sonics, Inc. Acoustic cell separation techniques and processes
US11021699B2 (en) 2015-04-29 2021-06-01 FioDesign Sonics, Inc. Separation using angled acoustic waves
US11474085B2 (en) 2015-07-28 2022-10-18 Flodesign Sonics, Inc. Expanded bed affinity selection
US11459540B2 (en) 2015-07-28 2022-10-04 Flodesign Sonics, Inc. Expanded bed affinity selection
US10709775B2 (en) * 2015-08-11 2020-07-14 Cellectis Cells for immunotherapy engineered for targeting CD38 antigen and for CD38 gene inactivation
MX2018005315A (es) 2015-10-30 2018-08-14 Aleta Biotherapeutics Inc Composiciones y metodos para el tratamiento del cancer.
MX2018005348A (es) 2015-10-30 2018-08-14 Aleta Biotherapeutics Inc Composiciones y metodos para transduccion de tumores.
CN105331586B (zh) * 2015-11-20 2020-09-15 上海细胞治疗研究院 一种包含高效杀伤启动机制的肿瘤精准t细胞及其用途
CN108290955A (zh) * 2015-11-24 2018-07-17 塞勒兰特治疗公司 人源化抗-cll-1抗体
BR112018011089A2 (pt) 2015-12-04 2018-12-04 Intellia Therapeutics Inc composições e métodos para a imuno-oncologia
EP3202783A1 (en) * 2016-02-02 2017-08-09 Ecole Polytechnique Fédérale de Lausanne (EPFL) Engineered antigen presenting cells and uses thereof
US11365237B2 (en) * 2016-03-23 2022-06-21 Helmholtz Zentrum Muenchen—Deutsches Forschungszentrum Fuer Gesundheit Und Umwelt (Gmbh) Fusion proteins of PD-1 and 4-1BB
US11104738B2 (en) 2016-04-04 2021-08-31 Hemogenyx Pharmaceuticals Llc Monoclonal antibodies to human FLT3/FLK2 receptor protein
CA3019581A1 (en) 2016-04-04 2017-10-12 Hemogenyx Llc Method of eliminating hematopoietic stem cells/hematopoietic progenitors (hsc/hp) in a patient using bi-specific antibodies
US11085035B2 (en) 2016-05-03 2021-08-10 Flodesign Sonics, Inc. Therapeutic cell washing, concentration, and separation utilizing acoustophoresis
US11214789B2 (en) 2016-05-03 2022-01-04 Flodesign Sonics, Inc. Concentration and washing of particles with acoustics
EP3468994A1 (en) 2016-06-08 2019-04-17 Intrexon Corporation Cd33 specific chimeric antigen receptors
US11466291B2 (en) 2016-07-06 2022-10-11 Cellectis Sequential gene editing in primary immune cells
KR20190038919A (ko) * 2016-08-19 2019-04-09 얀센 바이오테크 인코포레이티드 항-nkg2d 항체로 크론병을 치료하는 방법
US20190307799A1 (en) 2016-09-23 2019-10-10 The Regents Of The University Of Michigan Engineered lymphocytes
AU2017338846B2 (en) * 2016-10-04 2020-09-24 Precision Biosciences, Inc. Co-stimulatory domains for use in genetically-modified cells
JP2020513248A (ja) 2016-10-19 2020-05-14 フロデザイン ソニックス， インク．Ｆｌｏｄｅｓｉｇｎ Ｓｏｎｉｃｓ， Ｉｎｃ． 音響による親和性細胞抽出
WO2018073394A1 (en) 2016-10-19 2018-04-26 Cellectis Cell death inducing chimeric antigen receptors
WO2018073393A2 (en) 2016-10-19 2018-04-26 Cellectis Tal-effector nuclease (talen) -modified allogenic cells suitable for therapy
KR20250072712A (ko) 2016-12-02 2025-05-26 앤젤레스 테라퓨틱스, 인코포레이티드 합성 면역 수용체 및 이의 사용 방법
WO2018115189A1 (en) 2016-12-21 2018-06-28 Cellectis Stably enginereed proteasome inhibitor resistant immune cells for immunotherapy
EP4219556A3 (en) 2016-12-21 2023-10-11 The United States of America as represented by The Secretary Department of Health and Human Services Human monoclonal antibodies specific for flt3 and uses thereof
EP3567049A4 (en) 2016-12-28 2020-08-26 Green Cross Lab Cell Corporation CHEMERICAL ANTIGENIC RECEPTOR AND NATURAL KILLER CELLS EXPRESSING THE SAME
CN108250301A (zh) * 2016-12-29 2018-07-06 天津天锐生物科技有限公司 一种多靶点嵌合抗原受体
WO2018160768A1 (en) 2017-02-28 2018-09-07 Vor Biopharma, Inc. Compositions and methods for inhibition lineage specific proteins
JP2020513832A (ja) 2017-03-22 2020-05-21 ノバルティス アーゲー 免疫腫瘍学のための組成物および方法
EP3601346A1 (en) 2017-03-29 2020-02-05 H. Hoffnabb-La Roche Ag Bispecific antigen binding molecule for a costimulatory tnf receptor
EP4414380A3 (en) 2017-03-31 2024-12-04 Cellectis SA Universal anti-cd22 chimeric antigen receptor engineered immune cells
IL269209B2 (en) 2017-04-01 2023-11-01 Avm Biotechnology Llc Replacement of cytotoxic preconditioning before cellular immunotherapy
US11622977B2 (en) 2017-05-12 2023-04-11 Crispr Therapeutics Ag Materials and methods for engineering cells and uses thereof in immuno-oncology
US11166985B2 (en) 2017-05-12 2021-11-09 Crispr Therapeutics Ag Materials and methods for engineering cells and uses thereof in immuno-oncology
PE20241587A1 (es) 2017-06-02 2024-08-01 Pfizer Receptores de antigenos quimericos que se dirigen a flt3
AU2018281316B2 (en) 2017-06-07 2024-05-30 Precigen, Inc. Expression of novel cell tags
JP2020524512A (ja) * 2017-06-21 2020-08-20 アイセル・ジーン・セラピューティクス・エルエルシー キメラ抗体受容体（ＣＡＲｓ）の組成物およびその使用方法
WO2019002633A1 (en) 2017-06-30 2019-01-03 Cellectis CELLULAR IMMUNOTHERAPY FOR REPETITIVE ADMINISTRATION
EP3651772A4 (en) * 2017-07-09 2021-04-07 BioSight Ltd. ANTI-CANCER POLYTHERAPY
CA3070409A1 (en) * 2017-07-20 2019-01-24 H. Lee Moffitt Cancer Center And Research Institute, Inc. Compositions and methods for targeting cd33-expressing cancers
WO2019016360A1 (en) 2017-07-21 2019-01-24 Cellectis MODIFIED IMMUNE CELLS RESISTANT TO TUMOR MICRO-ENVIRONMENT
EP3658163A4 (en) * 2017-07-25 2021-08-04 Board Of Regents, The University Of Texas System IMPROVED ANTIGENIC CHEMICAL RECEPTORS AND THEIR USES
WO2019020733A1 (en) 2017-07-26 2019-01-31 Cellectis METHODS OF IMMUNE CELL SELECTION OF THE ANTIGEN-DEPENDENT CHIMERIC ANTIGENIC RECEPTOR (CAR)
MX2020004185A (es) * 2017-09-27 2021-01-08 Univ Southern California Nuevas plataformas para coestimulacion, nuevos diseños car y otras mejoras para terapia celular adoptiva.
WO2019072824A1 (en) 2017-10-09 2019-04-18 Cellectis IMPROVED ANTI-CD123 CAR IN UNIVERSAL MODIFIED IMMUNE T LYMPHOCYTES
US11649294B2 (en) 2017-11-14 2023-05-16 GC Cell Corporation Anti-HER2 antibody or antigen-binding fragment thereof, and chimeric antigen receptor comprising same
CN111655732B (zh) 2017-11-14 2023-09-12 Gc细胞治疗 抗her2抗体或其抗原结合片段及包含其的嵌合抗原受体
CN109837303A (zh) * 2017-11-25 2019-06-04 深圳宾德生物技术有限公司 一种敲除pd1的靶向cd317的嵌合抗原受体t细胞及其制备方法和应用
CN109836496A (zh) * 2017-11-25 2019-06-04 深圳宾德生物技术有限公司 一种靶向cd317的单链抗体、嵌合抗原受体t细胞及其制备方法和应用
WO2019106163A1 (en) 2017-12-01 2019-06-06 Cellectis Reprogramming of genetically engineered primary immune cells
KR20220066413A (ko) 2017-12-14 2022-05-24 프로디자인 소닉스, 인크. 음향 트랜스듀서 구동기 및 제어기
CN109608548A (zh) * 2017-12-29 2019-04-12 郑州大学第附属医院 基于人源cd20抗体的嵌合抗原受体、慢病毒表达载体及其应用
CN111770937A (zh) * 2017-12-29 2020-10-13 希望之城 中间位使能t细胞
WO2019129850A1 (en) 2017-12-29 2019-07-04 Cellectis Off-the-shelf engineered cells for therapy
AU2018396083B2 (en) 2017-12-29 2025-09-25 Cellectis Method for improving production of CAR T cells
BR112020013954A2 (pt) * 2018-01-09 2020-12-01 H. Lee Moffitt Cancer Center And Research Institute Inc. composições e métodos para alvejamento de cânceres que expressam clec12a
PE20211299A1 (es) * 2018-02-01 2021-07-20 Pfizer Receptores de antigeno quimericos dirigidos a cd70
TWI878213B (zh) 2018-02-01 2025-04-01 美商輝瑞大藥廠 對cd70具特異性抗體及其用途
JP7526101B2 (ja) 2018-03-02 2024-07-31 アロジーン セラピューティクス，インコーポレイテッド 誘導性キメラサイトカイン受容体
KR20200131867A (ko) * 2018-03-14 2020-11-24 더 유나이티드 스테이츠 오브 어메리카, 애즈 리프리젠티드 바이 더 세크러테리, 디파트먼트 오브 헬쓰 앤드 휴먼 서비씨즈 항-cd33 키메라 항원 수용체 및 이의 용도
EP3768315A4 (en) * 2018-03-21 2022-11-30 Dana Farber Cancer Institute, Inc. Fc variant compositions and methods of use thereof
CN108530536B (zh) * 2018-03-27 2021-08-13 刘爽 Cart-cd123及其制备与应用
CN108531457A (zh) * 2018-04-10 2018-09-14 杭州荣泽生物科技有限公司 一种Cas9/RNP敲除T细胞PD-1和LAG3基因及制备CAR-T细胞的方法
BR112020020919A2 (pt) 2018-04-13 2021-04-06 Sangamo Therapeutics France Receptor de antígeno quimérico específico para receptor de interleucina-23
IL278348B2 (en) 2018-05-11 2025-03-01 Crispr Therapeutics Ag Methods and preparations for treating cancer
EP3796935A4 (en) 2018-05-23 2022-03-16 The Jackson Laboratory ANTI-OX40 ANTIBODIES AND METHODS OF USE
EP3802619A1 (en) 2018-06-08 2021-04-14 F. Hoffmann-La Roche AG Peptidic linker with reduced post-translational modification
US20220403001A1 (en) 2018-06-12 2022-12-22 Obsidian Therapeutics, Inc. Pde5 derived regulatory constructs and methods of use in immunotherapy
WO2019246593A2 (en) * 2018-06-22 2019-12-26 Fred Hutchinson Cancer Research Center Compositions and methods to target cll-1 and cd123 for the treatment of acute myeloid leukemia and related disorders
US20210268028A1 (en) * 2018-07-02 2021-09-02 Cellectis Chimeric antigen receptors (car)-expressing cells and combination treatment for immunotherapy of patients with relapse refractory adverse genetic risk aml
US20210308171A1 (en) * 2018-08-07 2021-10-07 The Broad Institute, Inc. Methods for combinatorial screening and use of therapeutic targets thereof
GB201813178D0 (en) * 2018-08-13 2018-09-26 Autolus Ltd Cell
MX2021002415A (es) 2018-08-28 2021-09-21 Vor Biopharma Inc Celulas madre hematopoyeticas modificadas geneticamente y usos de las mismas.
CN109503716B (zh) * 2018-10-08 2021-04-27 浙江生研生物科技有限公司 一种双特异性嵌合抗原受体分子及其在肿瘤治疗上的应用
US12331320B2 (en) 2018-10-10 2025-06-17 The Research Foundation For The State University Of New York Genome edited cancer cell vaccines
JP2022505921A (ja) * 2018-10-26 2022-01-14 カファ セラピューティクス リミテッド Ｃｌｌ１を標的とする抗体およびその応用
EP3873500A4 (en) 2018-10-31 2023-01-11 Mayo Foundation for Medical Education and Research METHODS AND MATERIALS FOR THE TREATMENT OF CANCER
WO2020092736A1 (en) 2018-10-31 2020-05-07 Mayo Foundation For Medical Education And Research Methods and materials for treating cancer
BR112021010279A2 (pt) 2018-12-01 2021-08-24 Allogene Therapeutics, Inc. Receptores de antígeno quimérico que alvejam antígeno de maturação de células b e métodos de uso dos mesmos
MX2021007339A (es) 2018-12-18 2021-07-15 Boehringer Ingelheim Io Canada Inc Anticuerpos agonistas de flt3 y usos de estos.
EP3931208A1 (en) 2019-03-01 2022-01-05 Allogene Therapeutics, Inc. Chimeric cytokine receptors bearing a pd-1 ectodomain
TW202045547A (zh) 2019-03-01 2020-12-16 美商艾洛基因醫療公司 靶向ｄｌｌ３的嵌合抗原受體和結合劑
MY206523A (en) 2019-03-01 2024-12-19 Allogene Therapeutics Inc Constitutively active chimeric cytokine receptors
SG11202109057XA (en) 2019-03-05 2021-09-29 Nkarta Inc Cd19-directed chimeric antigen receptors and uses thereof in immunotherapy
WO2020183158A1 (en) * 2019-03-11 2020-09-17 Leucid Bio Ltd MUC1 PARALLEL CAR (pCAR) THERAPEUTIC AGENTS
EP3938402A4 (en) * 2019-03-15 2023-03-08 Rutgers, The State University of New Jersey CHIMERIC CD147 ANTIGEN RECEPTORS AND METHODS OF USE
JP7638881B2 (ja) 2019-03-21 2025-03-04 アロジーン セラピューティクス，インコーポレイテッド ＴＣＲαβ＋細胞の枯渇効率を向上させる方法
EP3947471A4 (en) * 2019-03-27 2023-03-15 The Trustees of The University of Pennsylvania TN-MUC1 CHIMERIC ANTIGEN RECEPTOR (CAR) T-LYMPHOCYTE THERAPY
WO2020210719A1 (en) 2019-04-10 2020-10-15 Elevatebio Management, Inc. Flt3-specific chimeric antigen receptors and methods of using the same
AU2020261411B2 (en) 2019-04-26 2025-10-02 Allogene Therapeutics, Inc. Methods of manufacturing allogeneic car T cells
CA3134308A1 (en) 2019-04-26 2020-10-29 Thomas Charles PERTEL Rituximab-resistant chimeric antigen receptors and uses thereof
EA202192975A1 (ru) 2019-04-30 2022-02-01 Криспр Терапьютикс Аг Аллогенная клеточная терапия b-клеточных злокачественных новообразований с применением генетически сконструированных т-клеток, нацеливающихся на cd19
AU2020295715A1 (en) * 2019-06-19 2022-02-17 Julius-Maximilians-Universität Würzburg Ultramodular IgG3-based spacer domain and multi-function site for implementation in chimeric antigen receptor design
CN112111013A (zh) * 2019-06-22 2020-12-22 南京北恒生物科技有限公司 一种靶向claudin 18.2的通用型嵌合抗原受体T细胞、构建方法及其用途
CA3146912A1 (en) * 2019-07-09 2021-01-14 Hemogenyx Pharmaceuticals Llc Method of eliminating hematopoietic stem cells/hematopoietic progenitors (hsc/hp) in a patient using bi-specific antibodies
CN114341193A (zh) 2019-08-30 2022-04-12 艾洛基治疗公司 包括tgfβ结合结构域的嵌合细胞因子受体
CN115103910A (zh) 2019-10-03 2022-09-23 工匠开发实验室公司 具有工程化的双引导核酸的crispr系统
EP3808766A1 (en) 2019-10-15 2021-04-21 Sangamo Therapeutics France Chimeric antigen receptor specific for interleukin-23 receptor
JP2023507525A (ja) 2019-12-23 2023-02-22 セレクティス 固形腫瘍の癌免疫療法のための新規メソテリン特異性キメラ抗原受容体（ｃａｒ）
TW202140533A (zh) 2020-01-16 2021-11-01 美商異基因治療有限公司 靶向B 細胞成熟抗原之嵌合抗原受體及γ 分泌酵素抑制劑之組合療法
IL295470A (en) 2020-02-24 2022-10-01 Allogene Therapeutics Inc Bcma car-t cells with enhanced activities
CA3170369A1 (en) 2020-03-05 2022-04-14 Michal Shahar Methods and compositions for treating cancer with immune cells
EP4114471A4 (en) * 2020-03-06 2024-12-11 Purdue Research Foundation METHODS, COMPOUNDS AND COMPOSITIONS FOR MODIFYING CAR-T CELL ACTIVITY
CN116194130A (zh) 2020-03-16 2023-05-30 南加利福尼亚大学 新颖的抗原结合结构域和包含这些抗原结合结构域的合成抗原受体
CA3169301A1 (en) * 2020-03-18 2021-09-23 Shyr Jiann Li Anti-il4 receptor antibodies for veterinary use
EP4125946A4 (en) * 2020-03-27 2024-04-24 Board of Regents, The University of Texas System Monoclonal antibodies targeting hsp70 and therapeutic uses thereof
CN115803435A (zh) 2020-05-06 2023-03-14 塞勒克提斯公司 用于在细胞基因组中靶向插入外源序列的方法
US20230227525A1 (en) * 2020-05-08 2023-07-20 Smt Bio Co., Ltd. Chimeric antigen receptor for treatment of cancer
GB202007842D0 (en) * 2020-05-26 2020-07-08 Quell Therapeutics Ltd Polypeptide useful in adoptive cell therapy
CN113735973B (zh) * 2020-05-28 2023-05-09 中国科学院微生物研究所 一种抗SIRPα抗体及其应用
WO2021241729A1 (ja) * 2020-05-29 2021-12-02 ブライトパス・バイオ株式会社 抗ｃｄ７３抗体およびその用途
WO2022003158A1 (en) 2020-07-03 2022-01-06 Cellectis S.A. Method for determining potency of chimeric antigen receptor expressing immune cells
WO2022020456A2 (en) 2020-07-21 2022-01-27 Allogene Therapeutics, Inc. Chimeric antigen receptors with enhanced signaling and activities and uses thereof
US20230248825A1 (en) 2020-07-24 2023-08-10 Cellectis S.A. T-cells expressing immune cell engagers in allogenic settings
JP2023537048A (ja) 2020-08-07 2023-08-30 ネオジン セラピューティクス ビー．ブイ． 遺伝子改変したｔ細胞の濃縮方法
CN112079927B (zh) * 2020-09-18 2021-12-28 西安桑尼赛尔生物医药有限公司 一种cd123结合蛋白、含其的car及其应用
WO2022061837A1 (en) * 2020-09-27 2022-03-31 Jiangsu Cell Tech Medical Research Institute Co., Ltd. Fibronectin extra domain b (edb) -specific car-t for cancer
CN113896795B (zh) * 2020-12-11 2022-07-15 广州百暨基因科技有限公司 抗cll1抗体及其应用
CN113234169B (zh) * 2020-12-11 2022-11-01 广州百暨基因科技有限公司 靶向cll1嵌合抗原受体及其应用
CN113248622B (zh) * 2020-12-11 2022-11-01 广州百暨基因科技有限公司 一种靶向cll1和nkg2d配体的双靶点嵌合抗原受体及其应用
AU2021401934A1 (en) 2020-12-14 2023-06-22 Allogene Therapeutics, Inc. Methods and reagents for characterizing car t cells for therapies
WO2022133169A1 (en) 2020-12-18 2022-06-23 Century Therapeutics, Inc. Chimeric antigen receptor systems with adaptable receptor specificity
IL303270A (en) 2020-12-21 2023-07-01 Allogene Therapeutics Inc The CD45–GATE vehicle activates a protease
JP7623497B2 (ja) 2020-12-31 2025-01-28 サノフイ ＮＫｐ４６およびＣＤ１２３に結合する多機能性ナチュラルキラー（ＮＫ）細胞エンゲージャ
CN112694532B (zh) * 2021-01-12 2023-04-18 倍而达药业（苏州）有限公司 抗Siglec-15的抗体或其抗原结合片段及应用
CN112661816B (zh) * 2021-01-13 2022-10-11 江西省人民医院 用于利妥昔单抗血药浓度检测的人工抗原及试剂盒
AU2022212090A1 (en) 2021-01-28 2023-07-06 Allogene Therapeutics, Inc. Methods for transducing immune cells
KR20230142470A (ko) 2021-01-29 2023-10-11 알로젠 테라퓨틱스 인코포레이티드 동종이계 세포 산물의 T 세포 인식을 완화하기 위한TAP2, NLRC5, β2m, TRAC, RFX5, RFXAP, 및 RFXANK 중 하나 이상의 녹다운 또는 녹아웃
WO2022182801A1 (en) 2021-02-25 2022-09-01 Artisan Development Labs, Inc. Compositions and methods for targeting, editing, or modifying genes
KR20240007179A (ko) * 2021-04-30 2024-01-16 셀렉티스 에스.에이. 고형 종양 암 면역 요법을 위한 새로운 항-muc1 car 및 유전자 편집된 면역세포
WO2022256448A2 (en) 2021-06-01 2022-12-08 Artisan Development Labs, Inc. Compositions and methods for targeting, editing, or modifying genes
WO2022266075A1 (en) 2021-06-14 2022-12-22 Caribou Biosciences, Inc. Methods and materials for treating cancer using car constructs with an intracellular domain comprising a stap domain and a kinase domain or a stap domain and a phosphatase domain
US20220409665A1 (en) 2021-06-15 2022-12-29 Allogene Therapeutics, Inc. Selective targeting of host cd70+ alloreactive cells to prolong allogeneic car t cell persistence
CN115477704B (zh) * 2021-06-16 2024-02-23 四川大学华西医院 一种基于lox1构建的嵌合抗原受体免疫细胞制备及其应用
CN115477705B (zh) * 2021-06-16 2024-02-23 四川大学华西医院 一种基于颗粒酶b构建的嵌合抗原受体免疫细胞制备及其应用
WO2022266538A2 (en) 2021-06-18 2022-12-22 Artisan Development Labs, Inc. Compositions and methods for targeting, editing or modifying human genes
WO2023020471A1 (en) * 2021-08-16 2023-02-23 Utc Therapeutics (Shanghai) Co., Ltd. Cd123-targetting antibodies and uses thereof in cancer therapies
MX2024002590A (es) 2021-09-01 2024-03-22 Springworks Therapeutics Inc Sintesis de nirogacestat.
CN118339187A (zh) * 2021-12-31 2024-07-12 上海宏成药业有限公司 抗pd-1抗体及其用途
EP4486881A1 (en) 2022-03-01 2025-01-08 Celyntra Therapeutics SA Composition and methods for transgene insertion
CA3244596A1 (en) 2022-03-29 2023-10-05 Allogene Therapeutics Inc Chimeric switch receivers for converting immunoactive signals into stimulation signals
US20250333515A1 (en) 2022-04-11 2025-10-30 Vor Biopharma Inc. Binding agents and methods of use thereof
IL317227A (en) 2022-05-27 2025-01-01 Sanofi Sa Combined natural killer (NK) cells bind to NKP46 and BCMA variants with FC engineering
CN119654406A (zh) 2022-07-29 2025-03-18 艾洛基治疗公司 具有降低的基因表达以减轻免疫细胞识别的经工程化的细胞
WO2024051751A1 (zh) * 2022-09-06 2024-03-14 河北森朗生物科技有限公司 抗cd123的纳米抗体、嵌合抗原受体及其应用
PE20252306A1 (es) * 2022-09-20 2025-09-22 Dana Farber Cancer Inst Inc Endocitosis mediada por receptores para su degradacion dirigida y suministro de agentes terapeuticos
CN118240086B (zh) * 2022-10-08 2025-05-09 东莞市朋志生物科技有限公司 抗纤维蛋白降解产物抗体、检测纤维蛋白降解产物的试剂和试剂盒
TW202440623A (zh) 2022-11-28 2024-10-16 美商艾洛基因醫療公司 靶向密連蛋白１８﹒２之嵌合抗原受體及結合劑以及其用途
WO2024121385A1 (en) 2022-12-09 2024-06-13 Cellectis S.A. Two-dose regimen in immunotherapy
WO2024238565A1 (en) 2023-05-15 2024-11-21 Vor Biopharma Inc. Egf-like module containing mucin-like hormone-like 2 (erm2) binding agents and methods of use thereof
CN117783521B (zh) * 2023-11-23 2025-03-04 中国农业科学院兰州兽医研究所 检测非洲猪瘟病毒p30蛋白抗体的竞争ELISA试剂盒
CN117964761B (zh) * 2023-12-21 2025-04-01 华润生物医药有限公司 抗trem2抗体及其用途
WO2025233825A1 (en) * 2024-05-06 2025-11-13 Janssen Pharmaceutica Nv Enrichment of cells expressing a bird linker

Citations (104)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4683195A (en) 1986-01-30 1987-07-28 Cetus Corporation Process for amplifying, detecting, and/or-cloning nucleic acid sequences
EP0239400A2 (en) 1986-03-27 1987-09-30 Medical Research Council Recombinant antibodies and methods for their production
WO1991009967A1 (en) 1989-12-21 1991-07-11 Celltech Limited Humanised antibodies
US5037743A (en) 1988-08-05 1991-08-06 Zymogenetics, Inc. BAR1 secretion signal
US5143830A (en) 1986-05-15 1992-09-01 Holland Ian B Process for the production of a polypeptide
EP0519596A1 (en) 1991-05-17 1992-12-23 Merck & Co. Inc. A method for reducing the immunogenicity of antibody variable domains
US5225539A (en) 1986-03-27 1993-07-06 Medical Research Council Recombinant altered antibodies and methods of making altered antibodies
WO1993017105A1 (en) 1992-02-19 1993-09-02 Scotgen Limited Altered antibodies, products and processes relating thereto
EP0592106A1 (en) 1992-09-09 1994-04-13 Immunogen Inc Resurfacing of rodent antibodies
WO1994024277A1 (en) 1993-04-13 1994-10-27 Sloan-Kettering Institute For Cancer Research Protection of human bone marrow from high dose antifolate therapy using mutated human dihydrofolate reductase dna
US5530101A (en) 1988-12-28 1996-06-25 Protein Design Labs, Inc. Humanized immunoglobulins
US5565332A (en) 1991-09-23 1996-10-15 Medical Research Council Production of chimeric antibodies - a combinatorial approach
US5766886A (en) 1991-12-13 1998-06-16 Xoma Corporation Modified antibody variable domains
US5858358A (en) 1992-04-07 1999-01-12 The United States Of America As Represented By The Secretary Of The Navy Methods for selectively stimulating proliferation of T cells
US6010613A (en) 1995-12-08 2000-01-04 Cyto Pulse Sciences, Inc. Method of treating materials with pulsed electrical fields
US6352694B1 (en) 1994-06-03 2002-03-05 Genetics Institute, Inc. Methods for inducing a population of T cells to proliferate using agents which recognize TCR/CD3 and ligands which stimulate an accessory molecule on the surface of the T cells
US6407213B1 (en) 1991-06-14 2002-06-18 Genentech, Inc. Method for making humanized antibodies
US6534055B1 (en) 1988-11-23 2003-03-18 Genetics Institute, Inc. Methods for selectively stimulating proliferation of T cells
US6642043B1 (en) 1996-03-12 2003-11-04 Sloan-Kettering Institute For Cancer Research Double mutants of dihydrofolate reductase and methods of using same
US6692964B1 (en) 1995-05-04 2004-02-17 The United States Of America As Represented By The Secretary Of The Navy Methods for transfecting T cells
US20040058414A1 (en) 1988-12-28 2004-03-25 Queen Cary L. Humanized immunoglobulins
WO2004043344A2 (en) 2002-11-07 2004-05-27 Immunogen, Inc. Anti-cd33 antibodies and method for treatment of acute myeloid leukemia using the same
US6797514B2 (en) 2000-02-24 2004-09-28 Xcyte Therapies, Inc. Simultaneous stimulation and concentration of cells
WO2004083379A2 (en) 2003-03-14 2004-09-30 Walters Richard E Large volume ex vivo electroporation method
WO2005000894A2 (en) 2003-06-25 2005-01-06 Crucell Holland B.V. Binding molecules for the treatment of myeloid cell malignancies
US20050042664A1 (en) 2003-08-22 2005-02-24 Medimmune, Inc. Humanization of antibodies
US20050048617A1 (en) 2003-08-18 2005-03-03 Medimmune, Inc. Humanization of antibodies
US6867041B2 (en) 2000-02-24 2005-03-15 Xcyte Therapies, Inc. Simultaneous stimulation and concentration of cells
US20050100543A1 (en) 2003-07-01 2005-05-12 Immunomedics, Inc. Multivalent carriers of bi-specific antibodies
US6905874B2 (en) 2000-02-24 2005-06-14 Xcyte Therapies, Inc. Simultaneous stimulation and concentration of cells
US6905680B2 (en) 1988-11-23 2005-06-14 Genetics Institute, Inc. Methods of treating HIV infected subjects
US20050175606A1 (en) 2001-04-11 2005-08-11 Hua-Liang Huang Cyclic single-chain trispecific antibody
WO2006020258A2 (en) 2004-07-17 2006-02-23 Imclone Systems Incorporated Novel tetravalent bispecific antibody
US20060121005A1 (en) 2000-02-24 2006-06-08 Xcyte Therapies, Inc. Activation and expansion of cells
US7067318B2 (en) 1995-06-07 2006-06-27 The Regents Of The University Of Michigan Methods for transfecting T cells
US20070014794A1 (en) 1995-03-01 2007-01-18 Genentech, Inc. Method for making heteromultimeric polypeptides
US7175843B2 (en) 1994-06-03 2007-02-13 Genetics Institute, Llc Methods for selectively stimulating proliferation of T cells
WO2007024715A2 (en) 2005-08-19 2007-03-01 Abbott Laboratories Dual variable domain immunoglobin and uses thereof
WO2008121420A1 (en) 2007-03-30 2008-10-09 Memorial Sloan-Kettering Cancer Center Constitutive expression of costimulatory ligands on adoptively transferred t lymphocytes
US7446179B2 (en) 2000-11-07 2008-11-04 City Of Hope CD19-specific chimeric T cell receptor
US7446190B2 (en) 2002-05-28 2008-11-04 Sloan-Kettering Institute For Cancer Research Nucleic acids encoding chimeric T cell receptors
US7514537B2 (en) 2001-04-30 2009-04-07 City Of Hope Chimeric immunoreceptor useful in treating human gliomas
WO2009051974A1 (en) 2007-10-17 2009-04-23 Nuvelo, Inc. Antibodes to cll-1
WO2009091826A2 (en) 2008-01-14 2009-07-23 The Board Of Regents Of The University Of Texas System Compositions and methods related to a human cd19-specific chimeric antigen receptor (h-car)
WO2010025177A1 (en) 2008-08-26 2010-03-04 City Of Hope Method and compositions for enhanced anti-tumor effector functioning of t cells
WO2011036183A2 (de) 2009-09-25 2011-03-31 Technische Universität Dresden Anti-cd33 antikörper und ihre anwendung zum immunotargeting bei der behandlung von cd33-assoziierten erkrankungen
EP2332994A1 (en) 2009-12-09 2011-06-15 Friedrich-Alexander-Universität Erlangen-Nürnberg Trispecific therapeutics against acute myeloid leukaemia
WO2012031744A1 (en) 2010-09-08 2012-03-15 Chemotherapeutisches Forschungsinstitut Chimeric antigen receptors with an optimized hinge region
WO2012050374A2 (en) 2010-10-13 2012-04-19 Innocell, Inc. Immunotherapy for solid tumors
WO2012058458A2 (en) 2010-10-27 2012-05-03 Cellectis Sa Method for increasing the efficiency of double-strand break-induced mutagenesis
WO2012079000A1 (en) 2010-12-09 2012-06-14 The Trustees Of The University Of Pennsylvania Use of chimeric antigen receptor-modified t cells to treat cancer
WO2012099973A2 (en) 2011-01-18 2012-07-26 The Trustees Of The University Of Pennsylvania Compositions and methods for treating cancer
WO2012136231A1 (en) 2010-09-08 2012-10-11 Chemotherapeutisches Forschungsinstitut Georg-Speyer-Haus Interleukin 15 as selectable marker for gene transfer in lymphocytes
WO2012138927A2 (en) 2011-04-05 2012-10-11 Philippe Duchateau Method for the generation of compact tale-nucleases and uses thereof
WO2012138475A1 (en) 2011-04-08 2012-10-11 The United States Of America, As Represented By The Secretary, Department Of Health And Human Services Anti-epidermal growth factor receptor variant iii chimeric antigen receptors and use of same for the treatment of cancer
US8324353B2 (en) 2001-04-30 2012-12-04 City Of Hope Chimeric immunoreceptor useful in treating human gliomas
US8399645B2 (en) 2003-11-05 2013-03-19 St. Jude Children's Research Hospital, Inc. Chimeric receptors with 4-1BB stimulatory signaling domain
WO2013040557A2 (en) 2011-09-16 2013-03-21 The Trustees Of The University Of Pennsylvania Rna engineered t cells for the treatment of cancer
WO2013059593A1 (en) 2011-10-20 2013-04-25 The United States Of America, As Represented By The Secretary, Department Of Health And Human Services Anti-cd22 chimeric antigen receptors
WO2013067492A1 (en) 2011-11-03 2013-05-10 The Trustees Of The University Of Pennsylvania Isolated b7-h4 specific compositions and methods of use thereof
WO2013070468A1 (en) 2011-11-08 2013-05-16 The Trustees Of The University Of Pennsylvania Glypican-3-specific antibody and uses thereof
WO2013074916A1 (en) 2011-11-18 2013-05-23 Board Of Regents, The University Of Texas System Car+ t cells genetically modified to eliminate expression of t- cell receptor and/or hla
WO2013123061A1 (en) 2012-02-13 2013-08-22 Seattle Children's Hospital D/B/A Seattle Children's Research Institute Bispecific chimeric antigen receptors and therapeutic uses thereof
WO2013126712A1 (en) 2012-02-22 2013-08-29 The Trustees Of The University Of Pennsylvania Compositions and methods for generating a persisting population of t cells useful for the treatment of cancer
WO2013126729A1 (en) 2012-02-22 2013-08-29 The Trustees Of The University Of Pennsylvania Use of the cd2 signaling domain in second-generation chimeric antigen receptors
WO2013126733A1 (en) 2012-02-22 2013-08-29 The Trustees Of University Of Pennsylvania Use of icos-based cars to enhance antitumor activity and car persistence
WO2013126726A1 (en) 2012-02-22 2013-08-29 The Trustees Of The University Of Pennsylvania Double transgenic t cells comprising a car and a tcr and their methods of use
US20130266551A1 (en) 2003-11-05 2013-10-10 St. Jude Children's Research Hospital, Inc. Chimeric receptors with 4-1bb stimulatory signaling domain
WO2013153391A1 (en) 2012-04-13 2013-10-17 Ucl Business Plc Polypeptide useful in adoptive cell therapy
CA2869562A1 (en) 2012-04-11 2013-10-17 The United States Of America, As Represented By The Secretary, Department Of Health And Human Services Chimeric antigen receptors targeting b-cell maturation antigen
US20130295118A1 (en) 2012-05-07 2013-11-07 Cellerant Therapeutics, Inc. Antibodies specific for cll-1
US20130309223A1 (en) 2012-05-18 2013-11-21 Seattle Genetics, Inc. CD33 Antibodies And Use Of Same To Treat Cancer
WO2013176915A1 (en) 2012-05-25 2013-11-28 Roman Galetto Methods for engineering allogeneic and immunosuppressive resistant t cell for immunotherapy
WO2014011987A1 (en) 2012-07-13 2014-01-16 The Trustees Of The University Of Pennsylvania Compositions and methods for regulating car t cells
WO2014011988A2 (en) 2012-07-13 2014-01-16 The Trustees Of The University Of Pennsylvania Enhancing activity of car t cells by co-introducing a bispecific antibody
US20140022163A1 (en) 2011-08-18 2014-01-23 Google Inc. Wearable device with input and output structures
US20140023674A1 (en) 2011-03-28 2014-01-23 St. Jude Children's Research Hospital Methods and compositions employing immunogenic fusion proteins
US20140024809A1 (en) 2011-02-11 2014-01-23 Memorial Sloan-Kettering Cancer Center Hla-restricted, peptide-specific antigen binding proteins
WO2014039523A1 (en) 2012-09-04 2014-03-13 Cellectis Multi-chain chimeric antigen receptor and uses thereof
WO2014100385A1 (en) 2012-12-20 2014-06-26 Anthrogenesis Corporation Chimeric antigen receptors
EP2765193A1 (en) 2011-10-07 2014-08-13 Mie University Chimeric antigen receptor
WO2014130635A1 (en) 2013-02-20 2014-08-28 Novartis Ag Effective targeting of primary human leukemia using anti-cd123 chimeric antigen receptor engineered t cells
WO2014145252A2 (en) 2013-03-15 2014-09-18 Milone Michael C Targeting cytotoxic cells with chimeric receptors for adoptive immunotherapy
WO2014144622A2 (en) 2013-03-15 2014-09-18 Stephen Forman Cd123-specific chimeric antigen receptor redirected t cells and methods of their use
WO2014153270A1 (en) 2013-03-16 2014-09-25 Novartis Ag Treatment of cancer using humanized anti-cd19 chimeric antigen receptor
WO2014152177A1 (en) 2013-03-15 2014-09-25 Anthrogenesis Corporation Modified t lymphocytes
WO2014184744A1 (en) 2013-05-13 2014-11-20 Cellectis Methods for engineering highly active t cell for immunotherapy
WO2014184143A1 (en) 2013-05-13 2014-11-20 Cellectis Cd19 specific chimeric antigen receptor and uses thereof
WO2014191128A1 (en) 2013-05-29 2014-12-04 Cellectis Methods for engineering t cells for immunotherapy by using rna-guided cas nuclease system
WO2015075195A1 (en) 2013-11-22 2015-05-28 Cellectis Method of engineering chemotherapy drug resistant t-cells for immunotherapy
WO2015075468A1 (en) 2013-11-21 2015-05-28 Ucl Business Plc Cell
WO2015121454A1 (en) 2014-02-14 2015-08-20 Cellectis Cells for immunotherapy engineered for targeting antigen present both on immune cells and pathological cells
WO2015123527A1 (en) 2014-02-14 2015-08-20 Bellicum Pharmaceuticals, Inc. Methods for activating t cells using an inducible chimeric polypeptide
WO2015136001A1 (en) 2014-03-11 2015-09-17 Cellectis Method for generating t-cells compatible for allogenic transplantation
WO2015142675A2 (en) 2014-03-15 2015-09-24 Novartis Ag Treatment of cancer using chimeric antigen receptor
WO2015140268A1 (en) 2014-03-19 2015-09-24 Cellectis Cd123 specific chimeric antigen receptors for cancer immunotherapy
WO2016014535A1 (en) 2014-07-21 2016-01-28 Novartis Ag Treatment of cancer using a cll-1 chimeric antigen receptor
US20160046724A1 (en) 2014-07-21 2016-02-18 The Trustees Of The University Of Pennsylvania Treatment of cancer using humanized anti-bcma chimeric antigen receptor
WO2016090369A1 (en) 2014-12-05 2016-06-09 City Of Hope Cs1 targeted chimeric antigen receptor-modified t cells
US20160184362A1 (en) 2013-05-29 2016-06-30 Cellectis Methods for engineering t cells for immunotherapy by using rna-guided cas nuclease system
US20160237139A1 (en) 2013-10-10 2016-08-18 Ucl Business Plc Chimeric antigen receptor
US20160297884A1 (en) 2015-04-13 2016-10-13 Pfizer Inc. Chimeric antigen receptors targeting b-cell maturation antigen
US20170073423A1 (en) 2013-12-20 2017-03-16 Cellectis Method of engineering multi-input signal sensitive t cell for immunotherapy
US20170105161A1 (en) 2015-10-12 2017-04-13 Telefonaktiebolaget Lm Ericsson (Publ) Cell boundary crossing in a unidirectional sfn for high speed trains

Family Cites Families (24)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
FR901228A (fr) 1943-01-16 1945-07-20 Deutsche Edelstahlwerke Ag Système d'aimant à entrefer annulaire
US4975278A (en) 1988-02-26 1990-12-04 Bristol-Myers Company Antibody-enzyme conjugates in combination with prodrugs for the delivery of cytotoxic agents to tumor cells
DE60138558D1 (de) 2000-09-13 2009-06-10 Multimmune Gmbh Peptid aus hsp70 welches nk zellaktivtät stimuliert und seine verwendung
RS20100366A (sr) * 2002-12-16 2011-04-30 Genentech, Inc. Varijante imunoglobulina i njihova upotreba
JP4898452B2 (ja) 2003-12-05 2012-03-14 マルチミューン ジーエムビーエイチ 治療および診断用抗Ｈｓｐ７０抗体
KR20060131892A (ko) * 2004-02-16 2006-12-20 마이크로메트 에이지 덜 면역원성인 결합 분자
GB0505971D0 (en) 2005-03-23 2005-04-27 Isis Innovation Delivery of molecules to a lipid bilayer
CN101316610A (zh) * 2005-09-22 2008-12-03 艾润·R·科恩 T细胞受体恒定区的免疫原性片段及其衍生的肽
WO2008102121A1 (en) 2007-02-20 2008-08-28 Oxford Nanopore Technologies Limited Formation of lipid bilayers
EP2072527A1 (en) * 2007-12-21 2009-06-24 Altonabiotec AG Fusion polypeptides comprising a SHBG dimerization component and uses thereof
US8956828B2 (en) * 2009-11-10 2015-02-17 Sangamo Biosciences, Inc. Targeted disruption of T cell receptor genes using engineered zinc finger protein nucleases
US20130022629A1 (en) * 2010-01-04 2013-01-24 Sharpe Arlene H Modulators of Immunoinhibitory Receptor PD-1, and Methods of Use Thereof
TWI580430B (zh) * 2010-10-27 2017-05-01 巴克斯歐塔公司 用於免疫耐受誘導及免疫診斷之ｆｖｉｉｉ胜肽
US8968539B2 (en) 2011-03-08 2015-03-03 Electronic Biosciences, Inc. Methods for voltage-induced protein incorporation into planar lipid bilayers
MX359513B (es) * 2011-03-23 2018-10-01 Hutchinson Fred Cancer Res Metodo y composiciones para inmunoterapia celular.
CA2852099C (en) * 2011-10-10 2020-12-08 City Of Hope Meditopes and meditope-binding antibodies and uses thereof
US9585969B2 (en) * 2011-11-15 2017-03-07 The Walter And Eliza Hall Institute Of Medical Research Soluble mediator
US20150017136A1 (en) 2013-07-15 2015-01-15 Cellectis Methods for engineering allogeneic and highly active t cell for immunotherapy
CN103483452B (zh) * 2012-06-12 2021-08-13 上海细胞治疗集团有限公司 双信号独立的嵌合抗原受体及其用途
WO2014151960A2 (en) 2013-03-14 2014-09-25 Bellicum Pharmaceuticals, Inc. Methods for controlling t cell proliferation
KR20230005422A (ko) * 2013-06-10 2023-01-09 다나-파버 캔서 인스티튜트 인크. 종양 세포에 의한 면역 억제를 감소시키기 위한 방법 및 조성물
PL3083671T3 (pl) * 2013-12-20 2021-03-22 Fred Hutchinson Cancer Center Znakowane chimeryczne cząsteczki efektorowe i ich receptory
JP6698546B2 (ja) * 2014-04-14 2020-05-27 セレクティスＣｅｌｌｅｃｔｉｓ 癌免疫療法のためのｂｃｍａ（ｃｄ２６９）特異的キメラ抗原受容体
EP2990416B1 (en) * 2014-08-29 2018-06-20 GEMoaB Monoclonals GmbH Universal chimeric antigen receptor expressing immune cells for targeting of diverse multiple antigens and method of manufacturing the same and use of the same for treatment of cancer, infections and autoimmune disorders

Patent Citations (119)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4683195B1 (enExample) 1986-01-30 1990-11-27 Cetus Corp
US4683195A (en) 1986-01-30 1987-07-28 Cetus Corporation Process for amplifying, detecting, and/or-cloning nucleic acid sequences
US5225539A (en) 1986-03-27 1993-07-06 Medical Research Council Recombinant altered antibodies and methods of making altered antibodies
EP0239400A2 (en) 1986-03-27 1987-09-30 Medical Research Council Recombinant antibodies and methods for their production
US5143830A (en) 1986-05-15 1992-09-01 Holland Ian B Process for the production of a polypeptide
US5037743A (en) 1988-08-05 1991-08-06 Zymogenetics, Inc. BAR1 secretion signal
US5883223A (en) 1988-11-23 1999-03-16 Gray; Gary S. CD9 antigen peptides and antibodies thereto
US6534055B1 (en) 1988-11-23 2003-03-18 Genetics Institute, Inc. Methods for selectively stimulating proliferation of T cells
US6905680B2 (en) 1988-11-23 2005-06-14 Genetics Institute, Inc. Methods of treating HIV infected subjects
US6887466B2 (en) 1988-11-23 2005-05-03 Genetics Institute, Inc. Methods for selectively stimulating proliferation of T cells
US7144575B2 (en) 1988-11-23 2006-12-05 The Regents Of The University Of Michigan Methods for selectively stimulating proliferation of T cells
US7232566B2 (en) 1988-11-23 2007-06-19 The United States As Represented By The Secretary Of The Navy Methods for treating HIV infected subjects
US5585089A (en) 1988-12-28 1996-12-17 Protein Design Labs, Inc. Humanized immunoglobulins
US5530101A (en) 1988-12-28 1996-06-25 Protein Design Labs, Inc. Humanized immunoglobulins
US20040058414A1 (en) 1988-12-28 2004-03-25 Queen Cary L. Humanized immunoglobulins
WO1991009967A1 (en) 1989-12-21 1991-07-11 Celltech Limited Humanised antibodies
EP0519596A1 (en) 1991-05-17 1992-12-23 Merck & Co. Inc. A method for reducing the immunogenicity of antibody variable domains
US6407213B1 (en) 1991-06-14 2002-06-18 Genentech, Inc. Method for making humanized antibodies
US5565332A (en) 1991-09-23 1996-10-15 Medical Research Council Production of chimeric antibodies - a combinatorial approach
US5766886A (en) 1991-12-13 1998-06-16 Xoma Corporation Modified antibody variable domains
WO1993017105A1 (en) 1992-02-19 1993-09-02 Scotgen Limited Altered antibodies, products and processes relating thereto
US5858358A (en) 1992-04-07 1999-01-12 The United States Of America As Represented By The Secretary Of The Navy Methods for selectively stimulating proliferation of T cells
EP0592106A1 (en) 1992-09-09 1994-04-13 Immunogen Inc Resurfacing of rodent antibodies
WO1994024277A1 (en) 1993-04-13 1994-10-27 Sloan-Kettering Institute For Cancer Research Protection of human bone marrow from high dose antifolate therapy using mutated human dihydrofolate reductase dna
US6352694B1 (en) 1994-06-03 2002-03-05 Genetics Institute, Inc. Methods for inducing a population of T cells to proliferate using agents which recognize TCR/CD3 and ligands which stimulate an accessory molecule on the surface of the T cells
US7175843B2 (en) 1994-06-03 2007-02-13 Genetics Institute, Llc Methods for selectively stimulating proliferation of T cells
US6905681B1 (en) 1994-06-03 2005-06-14 Genetics Institute, Inc. Methods for selectively stimulating proliferation of T cells
US20070014794A1 (en) 1995-03-01 2007-01-18 Genentech, Inc. Method for making heteromultimeric polypeptides
US6692964B1 (en) 1995-05-04 2004-02-17 The United States Of America As Represented By The Secretary Of The Navy Methods for transfecting T cells
US7172869B2 (en) 1995-05-04 2007-02-06 The United States Of America As Represented By The Secretary Of The Navy Methods for transfecting T cells
US7067318B2 (en) 1995-06-07 2006-06-27 The Regents Of The University Of Michigan Methods for transfecting T cells
US6010613A (en) 1995-12-08 2000-01-04 Cyto Pulse Sciences, Inc. Method of treating materials with pulsed electrical fields
US6642043B1 (en) 1996-03-12 2003-11-04 Sloan-Kettering Institute For Cancer Research Double mutants of dihydrofolate reductase and methods of using same
US6867041B2 (en) 2000-02-24 2005-03-15 Xcyte Therapies, Inc. Simultaneous stimulation and concentration of cells
US20060121005A1 (en) 2000-02-24 2006-06-08 Xcyte Therapies, Inc. Activation and expansion of cells
US6797514B2 (en) 2000-02-24 2004-09-28 Xcyte Therapies, Inc. Simultaneous stimulation and concentration of cells
US6905874B2 (en) 2000-02-24 2005-06-14 Xcyte Therapies, Inc. Simultaneous stimulation and concentration of cells
US7446179B2 (en) 2000-11-07 2008-11-04 City Of Hope CD19-specific chimeric T cell receptor
US20050175606A1 (en) 2001-04-11 2005-08-11 Hua-Liang Huang Cyclic single-chain trispecific antibody
US8324353B2 (en) 2001-04-30 2012-12-04 City Of Hope Chimeric immunoreceptor useful in treating human gliomas
US7514537B2 (en) 2001-04-30 2009-04-07 City Of Hope Chimeric immunoreceptor useful in treating human gliomas
US7446190B2 (en) 2002-05-28 2008-11-04 Sloan-Kettering Institute For Cancer Research Nucleic acids encoding chimeric T cell receptors
WO2004043344A2 (en) 2002-11-07 2004-05-27 Immunogen, Inc. Anti-cd33 antibodies and method for treatment of acute myeloid leukemia using the same
WO2004083379A2 (en) 2003-03-14 2004-09-30 Walters Richard E Large volume ex vivo electroporation method
WO2005000894A2 (en) 2003-06-25 2005-01-06 Crucell Holland B.V. Binding molecules for the treatment of myeloid cell malignancies
US20050100543A1 (en) 2003-07-01 2005-05-12 Immunomedics, Inc. Multivalent carriers of bi-specific antibodies
US20050048617A1 (en) 2003-08-18 2005-03-03 Medimmune, Inc. Humanization of antibodies
US20050042664A1 (en) 2003-08-22 2005-02-24 Medimmune, Inc. Humanization of antibodies
US8399645B2 (en) 2003-11-05 2013-03-19 St. Jude Children's Research Hospital, Inc. Chimeric receptors with 4-1BB stimulatory signaling domain
US20130266551A1 (en) 2003-11-05 2013-10-10 St. Jude Children's Research Hospital, Inc. Chimeric receptors with 4-1bb stimulatory signaling domain
WO2006020258A2 (en) 2004-07-17 2006-02-23 Imclone Systems Incorporated Novel tetravalent bispecific antibody
WO2007024715A2 (en) 2005-08-19 2007-03-01 Abbott Laboratories Dual variable domain immunoglobin and uses thereof
WO2008121420A1 (en) 2007-03-30 2008-10-09 Memorial Sloan-Kettering Cancer Center Constitutive expression of costimulatory ligands on adoptively transferred t lymphocytes
WO2009051974A1 (en) 2007-10-17 2009-04-23 Nuvelo, Inc. Antibodes to cll-1
WO2009091826A2 (en) 2008-01-14 2009-07-23 The Board Of Regents Of The University Of Texas System Compositions and methods related to a human cd19-specific chimeric antigen receptor (h-car)
WO2009091826A3 (en) 2008-01-14 2009-11-19 The Board Of Regents Of The University Of Texas System Compositions and methods related to a human cd19-specific chimeric antigen receptor (h-car)
WO2010025177A1 (en) 2008-08-26 2010-03-04 City Of Hope Method and compositions for enhanced anti-tumor effector functioning of t cells
WO2011036183A2 (de) 2009-09-25 2011-03-31 Technische Universität Dresden Anti-cd33 antikörper und ihre anwendung zum immunotargeting bei der behandlung von cd33-assoziierten erkrankungen
EP2332994A1 (en) 2009-12-09 2011-06-15 Friedrich-Alexander-Universität Erlangen-Nürnberg Trispecific therapeutics against acute myeloid leukaemia
WO2012031744A1 (en) 2010-09-08 2012-03-15 Chemotherapeutisches Forschungsinstitut Chimeric antigen receptors with an optimized hinge region
WO2012136231A1 (en) 2010-09-08 2012-10-11 Chemotherapeutisches Forschungsinstitut Georg-Speyer-Haus Interleukin 15 as selectable marker for gene transfer in lymphocytes
WO2012050374A2 (en) 2010-10-13 2012-04-19 Innocell, Inc. Immunotherapy for solid tumors
WO2012058458A2 (en) 2010-10-27 2012-05-03 Cellectis Sa Method for increasing the efficiency of double-strand break-induced mutagenesis
WO2012079000A1 (en) 2010-12-09 2012-06-14 The Trustees Of The University Of Pennsylvania Use of chimeric antigen receptor-modified t cells to treat cancer
WO2012099973A2 (en) 2011-01-18 2012-07-26 The Trustees Of The University Of Pennsylvania Compositions and methods for treating cancer
US20140024809A1 (en) 2011-02-11 2014-01-23 Memorial Sloan-Kettering Cancer Center Hla-restricted, peptide-specific antigen binding proteins
US20140023674A1 (en) 2011-03-28 2014-01-23 St. Jude Children's Research Hospital Methods and compositions employing immunogenic fusion proteins
WO2012138927A2 (en) 2011-04-05 2012-10-11 Philippe Duchateau Method for the generation of compact tale-nucleases and uses thereof
WO2012138475A1 (en) 2011-04-08 2012-10-11 The United States Of America, As Represented By The Secretary, Department Of Health And Human Services Anti-epidermal growth factor receptor variant iii chimeric antigen receptors and use of same for the treatment of cancer
US20140022163A1 (en) 2011-08-18 2014-01-23 Google Inc. Wearable device with input and output structures
WO2013040557A2 (en) 2011-09-16 2013-03-21 The Trustees Of The University Of Pennsylvania Rna engineered t cells for the treatment of cancer
EP2765193A1 (en) 2011-10-07 2014-08-13 Mie University Chimeric antigen receptor
WO2013059593A1 (en) 2011-10-20 2013-04-25 The United States Of America, As Represented By The Secretary, Department Of Health And Human Services Anti-cd22 chimeric antigen receptors
WO2013067492A1 (en) 2011-11-03 2013-05-10 The Trustees Of The University Of Pennsylvania Isolated b7-h4 specific compositions and methods of use thereof
WO2013070468A1 (en) 2011-11-08 2013-05-16 The Trustees Of The University Of Pennsylvania Glypican-3-specific antibody and uses thereof
WO2013074916A1 (en) 2011-11-18 2013-05-23 Board Of Regents, The University Of Texas System Car+ t cells genetically modified to eliminate expression of t- cell receptor and/or hla
WO2013123061A1 (en) 2012-02-13 2013-08-22 Seattle Children's Hospital D/B/A Seattle Children's Research Institute Bispecific chimeric antigen receptors and therapeutic uses thereof
WO2013126729A1 (en) 2012-02-22 2013-08-29 The Trustees Of The University Of Pennsylvania Use of the cd2 signaling domain in second-generation chimeric antigen receptors
WO2013126726A1 (en) 2012-02-22 2013-08-29 The Trustees Of The University Of Pennsylvania Double transgenic t cells comprising a car and a tcr and their methods of use
WO2013126733A1 (en) 2012-02-22 2013-08-29 The Trustees Of University Of Pennsylvania Use of icos-based cars to enhance antitumor activity and car persistence
WO2013126712A1 (en) 2012-02-22 2013-08-29 The Trustees Of The University Of Pennsylvania Compositions and methods for generating a persisting population of t cells useful for the treatment of cancer
CA2869562A1 (en) 2012-04-11 2013-10-17 The United States Of America, As Represented By The Secretary, Department Of Health And Human Services Chimeric antigen receptors targeting b-cell maturation antigen
WO2013154760A1 (en) 2012-04-11 2013-10-17 The United States Of America, As Represented By The Secretary, Department Of Health And Human Services Chimeric antigen receptors targeting b-cell maturation antigen
US9765342B2 (en) 2012-04-11 2017-09-19 The United States Of America, As Represented By The Secretary, Department Of Health And Human Services Chimeric antigen receptors targeting B-cell maturation antigen
WO2013153391A1 (en) 2012-04-13 2013-10-17 Ucl Business Plc Polypeptide useful in adoptive cell therapy
WO2013169625A1 (en) 2012-05-07 2013-11-14 Cellerant Therapeutics, Inc. Antibodies specific for cll-1
US20130295118A1 (en) 2012-05-07 2013-11-07 Cellerant Therapeutics, Inc. Antibodies specific for cll-1
US20130309223A1 (en) 2012-05-18 2013-11-21 Seattle Genetics, Inc. CD33 Antibodies And Use Of Same To Treat Cancer
WO2013176916A1 (en) 2012-05-25 2013-11-28 Roman Galetto Use of pre t alpha or functional variant thereof for expanding tcr alpha deficient t cells
WO2013176915A1 (en) 2012-05-25 2013-11-28 Roman Galetto Methods for engineering allogeneic and immunosuppressive resistant t cell for immunotherapy
US20140134142A1 (en) 2012-05-25 2014-05-15 Cellectis Multi-Chain Chimeric Antigen Receptor and Uses Thereof
WO2014011987A1 (en) 2012-07-13 2014-01-16 The Trustees Of The University Of Pennsylvania Compositions and methods for regulating car t cells
WO2014011988A2 (en) 2012-07-13 2014-01-16 The Trustees Of The University Of Pennsylvania Enhancing activity of car t cells by co-introducing a bispecific antibody
WO2014039523A1 (en) 2012-09-04 2014-03-13 Cellectis Multi-chain chimeric antigen receptor and uses thereof
WO2014100385A1 (en) 2012-12-20 2014-06-26 Anthrogenesis Corporation Chimeric antigen receptors
WO2014130635A1 (en) 2013-02-20 2014-08-28 Novartis Ag Effective targeting of primary human leukemia using anti-cd123 chimeric antigen receptor engineered t cells
WO2014145252A2 (en) 2013-03-15 2014-09-18 Milone Michael C Targeting cytotoxic cells with chimeric receptors for adoptive immunotherapy
WO2014144622A2 (en) 2013-03-15 2014-09-18 Stephen Forman Cd123-specific chimeric antigen receptor redirected t cells and methods of their use
WO2014152177A1 (en) 2013-03-15 2014-09-25 Anthrogenesis Corporation Modified t lymphocytes
US20140322183A1 (en) 2013-03-15 2014-10-30 The Trustees Of The University Of Pennsylvania Targeting cytotoxic cells with chimeric receptors for adoptive immunotherapy
WO2014153270A1 (en) 2013-03-16 2014-09-25 Novartis Ag Treatment of cancer using humanized anti-cd19 chimeric antigen receptor
WO2014184744A1 (en) 2013-05-13 2014-11-20 Cellectis Methods for engineering highly active t cell for immunotherapy
WO2014184143A1 (en) 2013-05-13 2014-11-20 Cellectis Cd19 specific chimeric antigen receptor and uses thereof
WO2014191128A1 (en) 2013-05-29 2014-12-04 Cellectis Methods for engineering t cells for immunotherapy by using rna-guided cas nuclease system
US20160184362A1 (en) 2013-05-29 2016-06-30 Cellectis Methods for engineering t cells for immunotherapy by using rna-guided cas nuclease system
US20160237139A1 (en) 2013-10-10 2016-08-18 Ucl Business Plc Chimeric antigen receptor
WO2015075468A1 (en) 2013-11-21 2015-05-28 Ucl Business Plc Cell
WO2015075195A1 (en) 2013-11-22 2015-05-28 Cellectis Method of engineering chemotherapy drug resistant t-cells for immunotherapy
US20170073423A1 (en) 2013-12-20 2017-03-16 Cellectis Method of engineering multi-input signal sensitive t cell for immunotherapy
WO2015123527A1 (en) 2014-02-14 2015-08-20 Bellicum Pharmaceuticals, Inc. Methods for activating t cells using an inducible chimeric polypeptide
WO2015121454A1 (en) 2014-02-14 2015-08-20 Cellectis Cells for immunotherapy engineered for targeting antigen present both on immune cells and pathological cells
WO2015136001A1 (en) 2014-03-11 2015-09-17 Cellectis Method for generating t-cells compatible for allogenic transplantation
WO2015142675A2 (en) 2014-03-15 2015-09-24 Novartis Ag Treatment of cancer using chimeric antigen receptor
WO2015140268A1 (en) 2014-03-19 2015-09-24 Cellectis Cd123 specific chimeric antigen receptors for cancer immunotherapy
WO2016014535A1 (en) 2014-07-21 2016-01-28 Novartis Ag Treatment of cancer using a cll-1 chimeric antigen receptor
US20160046724A1 (en) 2014-07-21 2016-02-18 The Trustees Of The University Of Pennsylvania Treatment of cancer using humanized anti-bcma chimeric antigen receptor
WO2016090369A1 (en) 2014-12-05 2016-06-09 City Of Hope Cs1 targeted chimeric antigen receptor-modified t cells
US20160297884A1 (en) 2015-04-13 2016-10-13 Pfizer Inc. Chimeric antigen receptors targeting b-cell maturation antigen
US20170105161A1 (en) 2015-10-12 2017-04-13 Telefonaktiebolaget Lm Ericsson (Publ) Cell boundary crossing in a unidirectional sfn for high speed trains

Non-Patent Citations (204)

* Cited by examiner, † Cited by third party
Title
"Treatment of Relapsed and/or Chemotherapy Refractory CD33 Positive Acute Myeloid Leukemia by CART-33 (CART33)," Feb. 1, 2015 [Online], May 20, 2013.
Arbiza, et al., "Characterization of two antigenic sites recognized by neutralizing monoclonal antibodies directed against the fusion glycoprotein of human respiratory syncytial virus." J Gen Viro. 1992; 73(9):2225-34).
Arimondo et al. "Exploring the cellular activity of camptothecin-triple-helix-forming oligonucleotide conjugates." Mol Cell Bioi 2006; 26(1): 324-33.
Atkins et al., "A case for "StopGo": Reprogramming translation to augment codon meaning of GGN by promoting unconventional termination (Stop) after addition of glycine and then allowing continued translation (Go)," RNA, 2007; 3: 803-810.
Ausubel et al., "Current Protocols in Molecular Biology," 2000, Wiley and Son Inc, Library of Congress, USA.
Baca, et al., "Antibody humanization using monovalent phage display." J Biol Chem. Apr. 18, 1997; 272(16):10678-84.
Bai, et al., "Involvement of miR-21 in resistance to daunorubicin by regulating PTEN expression in the leukaemia K562 cell line." FEBS Lett. Jan. 21, 2011; 585(2):402-8.
Bakker, et al., "C-type lectin-like molecule-1: a novel myeloid cell surface marker associated with acute myeloid leukemia", Cancer Res. 2004; 64(22):8443-50).
Bardenheuer, et al., "Resistance to cytarabine and gemcitabine and in vitro selection of transduced cells after retroviral expression of cytidine deaminase in human hematopoietic progenitor cells." Leukemia. Dec. 2005; 19(12):2281-8.
Barrow & Trowsdale, "You say ITAM and I say ITIM, let's call the whole thing off: the ambiguity of immunoreceptor signalling." Eur J Immunol. Jul. 2006; 36(7):1646-53.
Bauer; et al., "Activation of NK cells and T cells by NKG2D, a receptor for stress-inducible MICA." Science. Jul. 30, 1999; 285(5428):727-9.
Beckman, et al., "Antibody constructs in cancer therapy: protein engineering strategies to improve exposure in solid tumors." Cancer. Jan. 15, 2007; 109(2):170-9.
Behringer, et al., "Manipulating the Mouse Embryo", Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., 1986.
Betts, et al. "Sensitive and viable identification of antigenspecific CD8+ T cells by a flow cytometric assay for degranulation." J Immunol Methods 2003, 281(1-2): 65-78.
Bierer, et al., "Cyclosporin A and FK506: molecular mechanisms of immunosuppression and probes for transplantation biology," Curr Opin Immunol. 1993; 5(5):763-73.
Boch, et al., "Breaking the code of DNA binding specificity of TAL-type III effectors." Science 326(5959): 1509-12.
Brewin; et al. "Generation of EBV-specific cytotoxic T cells that are resistant to calcineurin inhibitors for the treatment of posttransplantation lymphoproliferative disease." Blood 114(23): 4792-803.
Bridgeman et al. (J. Immunol. Jun. 15, 2010; 184 (12): 6938-49). *
Byrd, et al., "Pretreatment cytogenetic abnormalities are predictive of induction success, cumulative incidence of relapse, and overall survival in adult patients with de novo acute myeloid leukemia: results from Cancer and Leukemia Group B (CALGB 8461)." Blood. Dec. 15, 2002; 100(13):4325-36.
Caldas et al. (Mol. Immunol. May 2003; 39 (15): 941-952). *
Carpenter, et al., "B-cell maturation antigen is a promising target for adoptive T-cell therapy of multiple myeloma." Clin Cancer Res. Apr. 15, 2013; 19(8):2048-60.
Cartellieri Marc et al., "A Novel Ex Vivo Isolation and Expansion Procedure for Chimeric Antigen Receptor Engrafted Human T Cells" Plos One, vol. 9, No. 4, Apr. 3, 2014; p. e93745.
Cartellieri, et al., "A novel ex vivo isolation and expansion procedure for chimeric antigen receptor engrafted human T cells." PLoS One. Apr. 3, 2014; 9(4):e93745.
Casset et al. (Biochem. Biophys. Res. Commun. Jul. 18, 2003; 307 (1): 198-205). *
Cespedes, "Mouse models in oncogenesis and cancer therapy." Clin Transl Oncol. May 2006; 8(5):318-29.
Chang et al. (Structure. Jan. 7, 2014; 22 (1): 9-21). *
Chen et al., "Molecular mechanisms of T cell co-stimulation and co-inhibition." Nat Rev Immunol. Apr. 2013; 13(4):227-42.
Chicaybam et al., "Abstract 2797: Construction and validation of an activating and inhibitory chimeric antigen receptor (CAR) system" Cancer Research 2014, 4 pages.
Chien et al. (Proc. Natl. Acad. Sci. USA. Jul. 1989; 86 (14): 5532-5536). *
Chien et al., "Significant structural and functional change of an antigen-binding site by a distant amino acid substitution: Proposal of a structural mechanism", Proc. Natl. Acad. Sci. USA 1989, 86:5532-5536.
Choulika, et al., "Induction of homologous recombination in mammalian chromosomes by using the 1-Scel system of Saccharomyces cerevisiae." Mol Cell Bio. 1995; 15(4): 1968-73.
Christian, et al. "Targeting DNA double-strand breaks with TAL effector nucleases." Genetics 2010; 186(2): 757-61.
Chu et al. "Genetic Modification of T Cells Redirected towards CS1 Enhances Eradication of Myeloma Cells", Clin. Cancer Res. Aug. 1, 2014; 20(15):3989-4000.
Coles, et al., "Increased CD200 expression in acute myeloid leukemia is linked with an increased frequency of FoxP3+ regulatory T cells." Leukemia. Sep. 2012; 26(9):2146-8.
Cong, et al., "Multiplex genome engineering using CRISPR/Cas systems." Science 2013; 339(6121): 819-23.
Couto, et al., "Anti-BA46 monoclonal antibody Mc3: humanization using a novel positional consensus and in vivo and in vitro characterization." Cancer Res. Apr. 15, 1995; 55(8):1717-22.
Couto, et al., "Designing human consensus antibodies with minimal positional templates." Cancer Res. Dec. 1, 1995;55(23 Suppl):5973s-5977s.
Dasgupta, et al., "Engineered drug-resistant immunocompetent cells enhance tumor cell killing during a chemotherapy challenge." Biochem Biophys Res Commun 2011; 391(1): 170-5.
Daugaard, "The heat shock protein 70 family: Highly homologous proteins with overlapping and distinct functions" FEBS Letters vol. 581, Issue 19, Jul. 31, 2007, pp. 3702-3710.
David C. Taussig, et al., Hematopoietic Stem Cells Express Multiple Myeloid Markers: Implications for the Origin and Targeted Therapy of Acute Myeloid Leukemia.
De Pascalis et al. (J. Immunol. 2002; 169 (6): 3076-3084). *
Deltcheva et al., "CRISPR RNA maturation by trans-encoded small RNA and host factor RNase III." Nature 471 2011; (7340): 602-7.
Deng, et al., "Structural basis for sequence-specific recognition of DNA by TAL effectors." Science 2012; 335(6069): 720-3.
Dennis, "Cancer: off by a whisker." Nature. Aug. 17, 2006; 442(7104):739-41.
Dixon, "Evaluation of the CASP2 docking section." Proteins. 1997; Suppl 1:198-204.
Domb, et al., "Focal Controlled Drug Delivery". Advances in Delivery Science and Technology. 2014.
Donnelly, et al., "Analysis of the aphthovirus 2A/2B polyprotein ‘cleavage’ mechanism indicates not a proteolytic eaction, but a novel translational effect: a putative ribosomal ‘skip’," J. of General Virology, 2001; 82: 1013-1025.
Donnelly, et al., "Nuclear localization and shuttling of herpes simplex virus tegument protein VP13/14." J Viral 2001; 75(6): 2566-74.
Donnelly, et al., "The cleavage activities of aphthovirus and cardiovirus 2A proteins.," J. of Gen. Virology, 1997; 78:13-21.
Doronina, et al., "Site-specific release of nascent chains from ribosomes at a sense codon." Mol Cell Bio 2008; 28(13}: 4227-39.
Dutour, et al., "In Vitro and In Vivo Antitumor Effect of Anti-CD33 Chimeric Receptor-Expressing EBV-CTL against CD33 Acute Myeloid Leukemia." Adv Hematol. 2012.
Eisenschmidt, et al., "Developing a programmed restriction endonuclease for highly specific DNA cleavage." Nucleic Acids Res 2005; 33(22): 7039-47.
Fedorov et al., "PD-1- and CTLA-4-based inhibitory chimeric antigen receptors (iCARs) divert off-target immunotherapy responses." Sci Transl Med. Dec. 11, 2013; 5(215):215ra172.
Finney, et al., "Activation of resting human primary T cells with chimeric receptors: costimulation from CD28, inducible costimulator, CD134, and CD137 in series with signals from the TCR zeta chain." J Immunol. Jan. 1, 2004; 172(1):104-13.
Freshney, et al, "Culture of Animal Cells," 1987, Wiley and Son Inc.
Fujimori, et al., "A modeling analysis of monoclonal antibody percolation through tumors: a binding-site barrier." J Nucl Med. Jul. 1990; 31(7):1191-8.
Gait, "Oligonucleotide Synthesis," 1984, IRL Press.
Garfall, et al., "Immunotherapy with chimeric antigen receptors for multiple myeloma." Discov Med. Jan. 2014; 17(91):37-46.
Garneau, et al., "The CRISPR/Cas bacterial immune system cleaves bacteriophage and plasmid DNA." Nature 2010; 468(7320): 67-71.
Gasiunas, et al., "Cas9-crRNA ribonucleoprotein complex mediates specific DNA cleavage for adaptive immunity in bacteria." Proc Natl Acad Sci U S A 2012; 109(39): E2579-86.
Gaudin, et al., "A hsp70-2 mutation recognized by CTL on a human renal cell carcinoma." J Immunol. 1999; 162(3):1730-8.
Geissler, et al., "Transcriptional activators of human genes with programmable DNA-specificity." PLoS One 2011; 6(5): e19509.
Giusti et al. (Proc. Natl. Acad. Sci. USA. May 1987; 84 (9): 2926-2930). *
Gorczynski, "CD200 and its receptors as targets for immunoregulation." Curr Opin Investig Drugs. May 2005; 6(5):483-8.
Gussow et al. (Methods in Enzymology. 1991; 203: 99-121). *
Hacke, et al., "Genetic modification of mouse bone marrow by lentiviral vector-mediated delivery of hypoxanthine-Guanine phosphoribosyltransferase short hairpin RNA confers chemoprotection against 6-thioguanine cytotoxicity." Transplant Proc 45(5): 2040-4.
Hames, et al., "Transcription and Translation" 1984, IRL Press.
Han, et al., "Chimeric antigen receptor-engineered T cells for cancer immunotherapy: progress and challenges." J Hematol Oncol. Jul. 8, 2013; 6:47.
Hanly, et al., "Review of Polyclonal Antibody Production Procedures in Mammals and Poultry." ILAR J. 1995; 37(3):93-118.
Hantschel, et al., "Hsp70 plasma membrane expression on primary tumor biopsy material and bone marrow of leukemic patients." Cell Stress Chaperones. 2000; 5(5):438-42).
Hau, et al., "Production of polyclonal antibodies: New technologies." ILAR J 2005; 46:294-299.
Heck, et al., "HSP70 expression: does it a novel fatigue signalling factor from immune system to the brain?" Cell Biochem Funct. 2011; 29(3):215-26.
Hellwig, et al., "TRAIL Signaling and Synergy Mechanisms Used in TRAIL-Based Combination Therapies", Mol Cancer Ther. 2012; 11(1):3-13.
Henderson, et al., "Comparison of the effects of FK-506, cyclosporin A and rapamycin on IL-2 production." Immunology 1991; 73(3): 316-21.
Holliger, et al., ""Diabodies": small bivalent and bispecific antibody fragments." 1993 Proc Natl Acad. Sci. U.S.A. 90:6444-6448.
Holm et al. (Mol. Immunol. Feb. 2007; 44 (6): 1075-1084). *
Hromadnikova, et al., "Expression of heat shock protein 70 and NKG2D ligands in acute myeloid leukemia cell lines." J Recept Signal Transduct Res. Jun. 2010; 30(3):161-9.
Hua Lu et al., Targeting Human C-Type Lectin-Like Molecule-1 (CLL1) With a Bispecific Antibody for Immunotherapy of Acute Myeloid Leukemia, 9841-9845; 2014; Angew. Chem. Int. Ed., Wiley-VCH Verlag GMBH & Co. KGAA, Weinheim.
Huang, et al., "Recombinant immunotherapeutics: current state and perspectives regarding the feasibility and market." Appl Microbiol Biotechnol. Jun. 2010; 87(2):401-10.
Hudecek et al. (Clin. Cancer Res. Jun. 15, 2013; 19 (12): 3153-64). *
Huston, et al., "Protein engineering of antibody binding sites: recovery of specific activity in an anti-digoxin single-chain Fv analogue produced in Escherichia coli." Proc Natl Acad Sci U S A. Aug. 1988; 85(16):5879-83.
International Search Report issued in International Application No. DK201570748 dated Dec. 18, 2015.
International Search Report issued in International Application No. PCT/EP2016/051469 dated Apr. 19, 2016.
Jakobsson, et al., "Identification and characterization of a novel human methyltransferase modulating Hsp70 function through lysine methylation."J. Bioi. Chem. 2013; 288:27752-27763.
Jena, et al., "Redirecting T-cell specificity by introducing a tumor-specific chimeric antigen receptor," Blood, 2010; 116 (7): 1035-1044.
Jensen, et al., "Design and implementation of adoptive therapy with chimeric antigen receptor-modified T cells." Immunol Rev. Jan. 2014; 257(1): 127-144.
Jinek, et al., "A programmable dual-RNA-guided DNA endonuclease in adaptive bacterial immunity." Science 2012; 337(6096): 816-21.
Jonnalagadda, et al., "Engineering human T cells for resistance to methotrexate and mycophenolate mofetil as an in vivo cell selection strategy." PLoS One 2013; 8(6): e65519.
Juillerat, et al., "An oxygen sensitive self-decision making engineered CAR T-cell." Sci Rep. Jan. 20, 2017; 7:39833.
Juillerat, et al., "Design of chimeric antigen receptors with integrated controllable transient functions." Sci Rep. Jan. 11, 2016; 6:18950.
Jushman, "Expression of human glutathione S-transferase P1 confers resistance to benzo[a]pyrene or benzo[a]pyrene-7,8-dihydrodiol mutagenesis, macromolecular alkylation and formation of stable N2-Gua-BPDE adducts in stably transfected V79MZ cells co-expressing hCYP1A1", Carcinogenesis vol. 28 No. 1 pp. 207-214, 2007.
Kabani, et al., "Multiple Hsp70 Isoforms in the Eukaryotic Cytosol: Mere Redundancy or Functional Specificity?" Curr Genomics. 2008; 9(5): 338-248.
Kevin J. Curran et al., Chimeric Antigen Receptors Fort Cell Immunotherapy: Current Understanding and Future Directions, pp. 405-415; 2012; John Wiley & Sons, Ltd, J Gene Med.
Kobayashi et al., "A chimeric antigen receptor for TRAIL-receptor 1 induces apoptosis in various types of tumor cells." Biochem Biophys Res Commun. Oct. 31, 2014;453(4):798-803.
Krause, et al., "The Chaperone Balance Hypothesis: The Importance of the Extracellular to Intracellular HSP70 Ratio to Inflammation-Driven Type 2 Diabetes, the Effect of Exercise, and the Implications for Clinical Management", Mediators of Inflammation 2015, vol. 2015, Article ID 249205, 12 pages.
Kushman, et al., "Expression of human glutathione S-transferase P1 confers resistance to benzo[a]pyrene or benzo[a]pyrene-7,8-dihydrodiol mutagenesis, macromolecular alkylation and formation of stable N2-Gua-BPDE adducts in stably transfected V79MZ cells co-expressing hCYP1A1." Carcinogenesis 2007; 28(1): 207-14.
Lamers et al., "Immune responses to transgene and retroviral vector in patients treated with ex vivo-engineered T cells", Blood, Jan. 6, 2011; 117(1):72-82.
Larsen, et al., "Expression of the hMICL in acute myeloid leukemia—a highly reliable disease marker at diagnosis and during follow-up". Cytometry B Clin Cytom. 2012; 82(1):3-8).
Lensink, "Docking and scoring protein complexes: CAPRI 3rd Edition." Proteins. Dec. 1, 2007; 69(4):704-18.
Lewandoski, "Conditional control of gene expression in the mouse." Nat Rev Genet. Oct. 2001; 2(10):743-55.
Li, et al., "Modularly assembled designer TAL effector nucleases for targeted gene knockout and gene replacement in eukaryotes." Nucleic Acids Res 2011; 39(14): 25 6315-25.
Li, et al., "Rapid and highly efficient construction of Tale-based transcriptional regulators and nucleases for genome modification." Plant Mol Bioi 2012; 78(4-5): 20 407-16.
Li, et al., "TAL nucleases (TALNs): hybrid proteins composed of TAL effectors and Fokl DNA-cleavage domain." Nucleic Acids Res 2011; 39(1): 359-72.
Liu, et al., "Calcineurin is a common target of cyclophilin-cyclosporin A and FKBP-FK506 complexes," Cell, 1991;66:807-815.
Liu, et al., "Inhibition of T cell signaling by immunophilin-ligand complexes correlates with loss of calcineurin phosphatase activity." Biochemistry 1992; 31(16): 3896-901.
Lu Hua et al.; "Targeting Human C-Type Lectin-like Molecule-1 (CLL1) with a Bispecific Antibody for Immunotherapy of Acute Myeloid Leukemia"; Angewandte Chemie International ED.; Sep. 8, 2014; vol. 53; pp. 9841-9845.
Ma, et al., "Yeast Mre11 and Rad1 proteins define a Ku-independent mechanism to repair double-strand breaks lacking overlapping end sequences." Mol Cell Bioi 2003; 23(23): 8820-8.
MacCallum et al. (J. Mol. Biol. Oct. 11, 1996; 262 (5): 732-745). *
Mahfouz, et al., "Targeted transcriptional repression using a chimeric TALE-SRDX repressor protein." Plant Mol Biol 2012; 78(3): 311-21.
Mak, et al., "The crystal structure of TAL effector PthXo1 bound to its DNA target." Science 2012; 335(6069): 716-9.
Mali, et al., "RNA-guided human genome engineering via Cas9." Science 2013; 339(6121): 823-6.
Malin, et al., "Enhanced metastasis suppression by targeting TRAIL receptor 2 in a murine model of triple-negative breast cancer." Clin Cancer Res. 2013; 17(15):5005-15.
Mardiros, et al., "T cells expressing CD123-specific chimeric antigen receptors exhibit specific cytolytic effector functions and antitumor effects against human acute myeloid leukemia," Blood, 2013; 122(18):3138-48.
Marin, et al., "Cytokine-induced killer cells for cell therapy of acute myeloid leukemia: improvement of their immune activity by expression of CD33-specific chimeric receptors." Haematologica. Dec. 2010;95(12):2144-52.
Mariuzza et al. (Annu. Rev. Biophys. Biophys. Chem. 1987; 16: 139-159). *
Marshall et al. (J. Biol. Chem. Apr. 9, 2004; 279 (15): 14792-802). *
Maus, et al., "Zoom Zoom: racing CARs for multiple myeloma." Clin Cancer Res. Apr. 15, 2013; 19(8):1917-9.
Mayer, et al., "Immunochemical Methods in Cell and Molecular Biology" 1987, Academic Press.
McGrath, "The tumor necrosis factor-related apoptosis-inducing ligand and lung cancer: still following the right TRAIL?" J Thorac Oncol. Jun. 2011;6(6):983-7.
Michael C. Jensen, et al., Design and Implementation of Adoptive Therapy With Chimeric Antigen Receptor-Modified T Cells, pp. 127-144, 2014, John Wiley & Sons Ltd., Immunol Rev.
Miller, et al., "A TALE nuclease architecture for efficient genome editing." Nat Biotechnol 2011; 29(2): 143-8.
Miller, et al., "Gene Transfer Vectors for Mammalian Cells" 1987, Cold Spring Harbor Laboratory.
Milone, et al., "Chimeric receptors containing CD137 signal transduction domains mediate enhanced survival of T cells and increased antileukemic efficacy in vivo." Mol Ther. Aug. 2009; 17(8):1453-64.
Morbitzer; et al., "Regulation of selected genome loci using de novoengineered transcription activator-like effector (TALE)-type transcription factors." Proc Natl Acad Sci USA 2011; 107(50): 21617-22.
Morrow, et al., "Multidrug resistance protein 1 (MRP1, ABCC1) mediates resistance to mitoxantrone via glutathione-dependent drug efflux." Mol Pharmacol. Apr. 2006; 69(4):1499-505.
Moscou, et al., "A simple cipher governs DNA recognition by TAL effectors." Science 2009; 326(5959): 1501.
Multhoff & Hightower, "Distinguishing integral and receptor-bound heat shock protein 70 (Hsp70) on the cell surface by Hsp70-specific antibodies." Cell Stress Chaperones. May 2011; 16(3):251-5.
Multhoff, "Heat shock protein 70 (Hsp70): membrane location, export and immunological relevance". Methods 2007; 43:229-237.
Mussolino, et al., "A novel TALE nuclease scaffold enables high genome editing activity in combination with low toxicity." Nucleic Acids Res 2011; 39(21): 9283-93.
Noorduis Paul et al., "Targeting of CLEC12A in Acute Myeloid Leukemia by Antibody-Drug-Conjugates and Bispecific CLL-1xCD3 BiTE Antibody"; blood; American Society of Hematology, Nov. 1, 2010; pp. 1-2.
Ohno et al., "Antigen binding specificities of antibodies are primarily determined by seven residues of VH", Proc. Natl. Acad. Sci. USA, 1985, 82(9): 2945-2949.
Ormhoj, et al., "CARs in the Lead Against Multiple Myeloma." Curr Hematol Malig Rep. Apr. 2017; 12(2):119-125.
Paques, et al., "Meganucleases and DNA double-strand break-induced recombination: perspectives for gene therapy." Curr Gene Ther 2007; 7(1): 49-66.
Park, et al., "Treating Cancer with Genetically Engineered T Cells," Trends Biotechnol., 2011; 29(11): 550-557.
Perbai "A Practical Guide to Molecular Cloning," 1984, Wiley & Sons.
Perrin, et al., "Asymmetrical recognition and activity of the 1-Scel endonuclease on its site and on intron-exon junctions." Embo J 1993; 12(7): 2939-47.
Philip et al., "A highly compact epitope-based marker/suicide gene for easier and safer T-Cell Therapy." Blood, Aug. 21, 2014, 124(8): 1277-1287.
Pingoud, et al. "Precision genome surgery." Nat Biotechnol 2007; 25(7): 743-4.
Pitti , et al., "Induction of apoptosis by Apo-2 ligand, a new member of the tumor necrosis factor cytokine family". J Bioi Chem 1996; 271:12687-90.
Pizzitola et al., "Chimeric antigen receptors against CD33/CD123 antigens efficiently target primary acute myeloid eukemia cells in vivo," Leukemia, 2014; 28(8):1596-605.
Pockley, et al., "Detection of heat shock protein 70 (Hsp70) and anti-Hsp70 antibodies in the serum of normal individuals." Immunol. Invest. 1998; 27 367-377.
Poirot, et al., "Multiplex Genome-Edited T-cell Manufacturing Platform for "Off-the-Shelf" Adoptive T-cell Immunotherapies." Cancer Res. Sep. 15, 2015; 75(18):3853-64.
Porter, et al., "Chimeric Antigen Receptor-Modified T Cells in Chronic Lymphoid Leukemia," N. Engl. J. Med., 2011; 365:725-733.
Porteus, et al., "Gene targeting using zinc finger nucleases." Nat Biotechnol 2005; 23(8): 967-73.
Quintarelli, "Co-expression of cytokine and suicide genes to enhance the activity and safety of tumor-specific cytotoxic T lymphocytes." Blood. Oct. 15, 2007; 110(8):2793-802.
Ravetch, et al., "Alternative membrane forms of Fe gamma RIii(CD16) on human natural killer cells and neutrophils. Cell type-specific expression of two genes that differ in single nucleotide substitutions". J. Exp. Med. 1989; 170:481-497.
Riechmann, et al., "Reshaping human antibodies for therapy." Nature. Mar. 24, 1988; 332(6162):323-7.
Riemer, et al., Vaccination with cetuximab mimotopes and biological properties of induced anti-epidermal growth factor receptor antibodies, J Natl Cancer Inst. 2005; 97(22):1663-70.
Roguska, et al., "A comparison of two murine monoclonal antibodies humanized by CDR-grafting and variable domain resurfacing." Protein Eng. Oct. 1996; 9(10):895-904.
Roguska, et al., "Humanization of murine monoclonal antibodies through variable domain resurfacing." Proc Natl Acad Sci U S A. Feb. 1, 1994;91(3):969-73.
Rouet, et al., "Introduction of double-strand breaks into the genome of mouse cells by expression of a rare-cutting endonuclease." Mol Cell Bio 1994; 14(12):8096-106.
Rudikoff et al. (Proc. Natl. Acad. Sci. USA. 1982; 79: 1979-1983). *
Rudikoff, et al., "Single amino acid substitution altering antigen-binding specificity", Proc. Nat. Acad. Sci. USA. Immunology, 1982, 79: abstract on p. 1979.
Rudnick, et al., "Affinity and avidity in antibody-based tumor targeting." Cancer Biother Radiopharm. Apr. 2009; 24(2):155-61.
Sadelain et al., The Basic Principles of Chimeric Antigen Receptor Design, pp. 388-398; 2013; Cancer Discovery; American Association for Cancer Research.
Sadelain Michel et al., "The Basic Principles of Chimeric Antigen Receptor Design" Cancer Discovery, vol. 3, No. 4, Apr. 1, 2013, pp. 388-398.
Sadelain, et al., "The promise and potential pitfalls of chimeric antigen receptors," Curr Opin Immunol, 2009; 21(2):215-23.
Sambrook, et al., "Molecular Cloning: A Laboratory Manual, Third Edition," 2001, Cold Spring Harbor, New York: Cold Spring Harbor Laboratory Press.
Sander, et al., "Targeted gene disruption in somatic zebrafish cells using engineered TALENs." Nat Biotechnol 2011; 29(8): 697-8.
Sangiolo, et al., "Lentiviral vector conferring resistance to mycophenolate mofetil and sensitivity to ganciclovir for in vivo T-cell selection." Gene Ther 2007; 14(21): 1549-54.
Scharfenberger, et al., "Transgenic mouse technology in skin biology: generation of complete or tissue-specific knockout mice." J Invest Dermatol. Jan. 2014; 134(1):1-5.
Schweitzer, et al., "Dihydrofolate reductase as a therapeutic target." Faseb J 1990; 4(8): 2441-52.
Schwenk, et al., "Temporally and spatially regulated somatic mutagenesis in mice." Nucleic Acids Res. Mar. 15, 1998; 26(6):1427-32.
Singer & Berg P., "Molecules of the Genetic Apparatus." Genes and Genomes, 1998, vol. 1, p. 63.
Smyth, et al., 2001, "Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) contributes to interferon gamma-dependent natural killer cell protection from tumor metastasis." J Exp Med;193:661-70.
Sorek, et al., "CRISPR-mediated Adaptive Immune Systems in Bacteria and Archaea." Annu Rev Biochem. 2013.
Stangl, et al., "Targeting membrane heat-shock protein 70 (Hsp70) on tumors by cmHsp70.1 antibody." Proc Natl Acad Sci U S A. Jan. 11, 2011;108(2):733-8.
Steiner, et al., "High HSP70-membrane expression on leukemic cells from patients with acute myeloid leukemia is associated with a worse prognosis." Leukemia. Nov. 2006; 20(11):2076-9.
Stoddard, "Homing endonuclease structure and function." Q Rev Biophys 2005; 38(1): 49-95.
Takebe, et al., "Generation of dual resistance to 4-hydroperoxycyclophosphamide and methotrexate by retroviral transfer of the human aldehyde dehydrogenase class 1 gene and a mutated dihydrofolate reductase gene." Mol Ther 2001; 3(1): 88-96.
Talmadge, et al., "Murine models to evaluate novel and conventional therapeutic strategies for cancer." Am J Pathol. Mar. 2007; 170(3):793-804.
Tame, et al., "Scoring functions: a view from the bench." J Comput Aided Mol Des. Mar. 1999; 13(2):99-108.
Tan, et al., ""Superhumanized" antibodies: reduction of immunogenic potential by complementarity-determining region grafting with human germline sequences: application to an anti-CD28."J Immunol. Jul. 15, 2002; 169(2):1119-25.
Tesson, et al. "Knockout rats generated by embryo microinjection of TALENs." Nat Biotechnol 2011; 29(8): 695-6.
Thaventhiran, "T cell co-inhibitory receptors-functions and signalling mechanisms." J. Clinical & Cellular Immunology 2012, 12 pages.
Thurber, et al., "Antibody tumor penetration: transport opposed by systemic and antigen-mediated clearance." Adv Drug Deliv Rev. Sep. 2008; 60(12):1421-34.
Vajdos et al. (J. Mol. Biol. Jul. 5, 2002; 320 (2): 415-428). *
Valton, et al., "A Multidrug-resistant Engineered CAR T Cell for Allogeneic Combination Immunotherapy." Mol Ther. Sep. 2015;23(9):1507-18.
Van Rhenen; et al., "The novel AML stem cell associated antigen CLL-1 aids in discrimination between normal and leukemic stem cells." Blood 2007; 110(7):2659-66.
Voskoglou-Nomikos, et al., "Clinical predictive value of the in vitro cell line, human xenograft, and mouse allograft preclinical cancer models." Clin Cancer Res. Sep. 15, 2003; 9(11):4227-39.
Wang, et al., "Treatment of CD33-directed chimeric antigen receptor-modified T cells in one patient with relapsed and refractory acute myeloid leukemia." Mol Ther. Jan. 2015; 23(1):184-91.
Weber, et al., "Assembly of designer TAL effectors by Golden Gate cloning." PLoS One 2011; 6(5): e19722.
Weir, et al., "Handbook of Experimental Immunology, vols. I-IV," 1986 Blackwell Scientific Publications.
White, "Membrane Protein Insertion: The Biology-Physics Nexus", J Gen Physiol. 2007; 129(5): 363-369.
Wiley; et al. "Identification and characterization of a new member of the TN F family that induces apoptosis." Immunity 1995;3:673-82.
Winkler et al. (J. Immunol. Oct. 15, 2000; 165 (8): 4505-4514). *
Woodward, "Immobilized Cells and Enzymes," 1986, IRL Press.
Wright; et al., "Characterization of the CD200 receptor family in mice and humans and their interactions with CD200." J Immunol. 2003; 171(6):3034-46.
Wu et al. (J. Mol. Biol. Nov. 19, 1999; 294 (1): 151-162). *
Wu, et al., "An activating immunoreceptor complex formed by NKG2D and DAP10." Science. Jul. 30, 1999; 285(5428):730-2.
Wu, et al., "Methods in Enzymology vol. 154. Recombinant DNA, Part E", 1987 Academic Press.
Wu, et al., "Methods in Enzymology vol. 155. Recombinant DNA, Part F" 1987 Academic Press.
Yam, et al., "Ex vivo selection and expansion of cells based on expression of a mutated inosine monophosphate dehydrogenase 2 after HIV vector transduction: effects on lymphocytes, monocytes, and CD34+ stem cells." Mol Ther 2006; 14(2): 15 236-44.
Yarilin A.A., "Molecular and cellular basis of adaptive immunity." Immunology Basics: Manual.—M.: Medicina, 1999, pp. 169-174.
Yarilin, A.A. "Osnovy immunologii", M: Medizina, 1999, p. 172-174.
Yu et al. (PLoS One. 2012; 7 (3): e33340; pp. 1-15). *
Zettlitz, et al., "Humanization of a mouse monoclonal antibody directed against a cell surface-exposed epitope of membrane-associated heat shock protein 70 (Hsp70)." REMol Biotechnol. 2010; 46(3):265-78).
Zhang, et al., "Characterization of high-affinity peptides and their feasibility for use in nanotherapeutics targeting leukemia stem cells" Nanomedicine 2011; 8(7): 1116-1124.
Zhang, et al., "Efficient construction of sequence-specific TAL effectors for modulating mammalian transcription." Nat Biotechnol 2011; 29(2): 149-53.
Zhang, et al., "Plasma levels of Hsp70 and anti-Hsp70 antibody predict risk of acute coronary syndrome" Cell Stress Chaperones. 2010; 15(5): 675-686.
Zhao Xiaoxian et al.; "Targeting C-type lectin-like molecule-1 for antibody-mediated immunotherapy in acute myeloid leukemia"; Haematologica, vol. 95, No. 1, Jul. 31, 2009; pp. 71-78.
Zhao, et al., "Targeting C-type lectin-like molecule-1 for antibody-mediated immunotherapy in acute myeloid leukemia." Haematologica. Jan. 2010; 95(1):71-8.
Zhuang Xiaodong et al.; "Abstract LB-256: Immunotherapy using genetically modified T lymphocytes to target CLEC14A on the tumor vasculature" Proceedings: AACR Annual Meeting 2014; Oct. 1, 2014, doi: 10,1158/1538-7445.AM2014-LB-256.
Zielske, et al., "In vivo selection of MGMT(P140K) lentivirustransduced human NOD/SCID repopulating cells without pretransplant irradiation conditioning. "J Clin Invest 2003; 112(10)1561-70.

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US12054531B2 (en) 2020-03-20 2024-08-06 Lyell Immunopharma, Inc. Recombinant cell surface markers
US12144827B2 (en) 2021-02-25 2024-11-19 Lyell Immunopharma, Inc. ROR1 targeting chimeric antigen receptor

Also Published As

Publication number Publication date
RU2017129592A (ru) 2019-02-28
EP3250607A1 (en) 2017-12-06
JP2018504143A (ja) 2018-02-15
RU2732925C2 (ru) 2020-09-24
RU2017129455A (ru) 2019-03-04
AU2016212162A1 (en) 2017-07-13
EP3250604A1 (en) 2017-12-06
JP6779909B2 (ja) 2020-11-04
US20180002427A1 (en) 2018-01-04
KR20170135822A (ko) 2017-12-08
WO2016120220A1 (en) 2016-08-04
RU2017129592A3 (enExample) 2019-07-17
CN107531801A (zh) 2018-01-02
US20180051089A1 (en) 2018-02-22
MX2017009181A (es) 2017-11-22
CN107406516A (zh) 2017-11-28
CA2973525C (en) 2021-12-14
ES2869972T3 (es) 2021-10-26
BR112017015453A2 (pt) 2018-01-23
CN113968914A (zh) 2022-01-25
AU2016212160A1 (en) 2017-07-13
CA2973532A1 (en) 2016-08-04
WO2016120219A1 (en) 2016-08-04
RU2017129591A (ru) 2019-03-04
US20240228636A1 (en) 2024-07-11
MX2017009182A (es) 2018-04-20
AU2016212160B2 (en) 2021-04-15
DK3250604T3 (da) 2021-07-12
JP6884709B2 (ja) 2021-06-09
IL252937A0 (en) 2017-08-31
CN107531801B (zh) 2021-11-12
JP2018504458A (ja) 2018-02-15
EP3250604B1 (en) 2021-04-21
KR20170134331A (ko) 2017-12-06
CA2973524A1 (en) 2016-08-04
KR102661969B1 (ko) 2024-04-29
CN107406516B (zh) 2021-11-23
JP2018504144A (ja) 2018-02-15
CA2973525A1 (en) 2016-08-04
EP3250606A1 (en) 2017-12-06
IL252937B (en) 2021-09-30
IL253049B (en) 2022-01-01
WO2016120216A1 (en) 2016-08-04
IL253049A0 (en) 2017-08-31
CN113968915A (zh) 2022-01-25
WO2016120217A1 (en) 2016-08-04
CA2973642A1 (en) 2016-08-04
DK3250606T3 (da) 2021-01-04
JP2018504145A (ja) 2018-02-15
ES2842212T3 (es) 2021-07-13
MX381908B (es) 2025-03-13
AU2016212159A1 (en) 2017-07-13
US20180009895A1 (en) 2018-01-11
AU2016212158A1 (en) 2017-07-13
RU2017129455A3 (enExample) 2019-08-15
US20180002435A1 (en) 2018-01-04
US20180000914A1 (en) 2018-01-04
CN107438618A (zh) 2017-12-05
MX389057B (es) 2025-03-20
JP2018504459A (ja) 2018-02-15
EP3250602A1 (en) 2017-12-06
RU2017129591A3 (enExample) 2019-08-05
KR102329836B1 (ko) 2021-11-19
WO2016120218A1 (en) 2016-08-04
CA2973529A1 (en) 2016-08-04
AU2016212161A1 (en) 2017-07-13
AU2016212158B2 (en) 2021-06-03
BR112017014547A2 (pt) 2018-01-16
IL253012A0 (en) 2017-08-31
EP3250606B1 (en) 2020-11-25
EP3250605A1 (en) 2017-12-06
MX2017009349A (es) 2017-09-26
RU2731543C2 (ru) 2020-09-04
KR20170135824A (ko) 2017-12-08
BR112017013981A2 (pt) 2018-01-02

Similar Documents

Publication Publication Date Title
US11014989B2 (en) Anti-CLL1 specific single-chain chimeric antigen receptors (scCARs) for cancer immunotherapy
US11919961B2 (en) CD123 specific chimeric antigen receptors for cancer immunotherapy
US11498971B2 (en) BCMA (CD269) specific chimeric antigen receptors for cancer immunotherapy

Legal Events

Date Code Title Description
STPP Information on status: patent application and granting procedure in general

Free format text: RESPONSE TO NON-FINAL OFFICE ACTION ENTERED AND FORWARDED TO EXAMINER

STPP Information on status: patent application and granting procedure in general

Free format text: NON FINAL ACTION MAILED

AS Assignment

Owner name: CELLECTIS, FRANCE

Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNORS:SMITH, JULIANNE;VALTON, JULIEN;DUCHATEAU, PHILIPPE;AND OTHERS;SIGNING DATES FROM 20181231 TO 20190128;REEL/FRAME:050135/0668

Owner name: RINAT NEUROSCIENCE CORP., CALIFORNIA

Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNORS:SASU, BARBRA JOHNSON;RAJPAL, ARVIND;SIGNING DATES FROM 20160518 TO 20160606;REEL/FRAME:050135/0805

STPP Information on status: patent application and granting procedure in general

Free format text: RESPONSE TO NON-FINAL OFFICE ACTION ENTERED AND FORWARDED TO EXAMINER

AS Assignment

Owner name: CELLECTIS, FRANCE

Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNOR:RINAT NEUROSCIENCE CORP.;REEL/FRAME:050215/0154

Effective date: 20160609

STPP Information on status: patent application and granting procedure in general

Free format text: FINAL REJECTION MAILED

STPP Information on status: patent application and granting procedure in general

Free format text: ADVISORY ACTION MAILED

STPP Information on status: patent application and granting procedure in general

Free format text: DOCKETED NEW CASE - READY FOR EXAMINATION

STPP Information on status: patent application and granting procedure in general

Free format text: RESPONSE TO NON-FINAL OFFICE ACTION ENTERED AND FORWARDED TO EXAMINER

STPP Information on status: patent application and granting procedure in general

Free format text: NOTICE OF ALLOWANCE MAILED -- APPLICATION RECEIVED IN OFFICE OF PUBLICATIONS

STPP Information on status: patent application and granting procedure in general

Free format text: PUBLICATIONS -- ISSUE FEE PAYMENT RECEIVED

STPP Information on status: patent application and granting procedure in general

Free format text: PUBLICATIONS -- ISSUE FEE PAYMENT VERIFIED

STCF Information on status: patent grant

Free format text: PATENTED CASE

MAFP Maintenance fee payment

Free format text: PAYMENT OF MAINTENANCE FEE, 4TH YEAR, LARGE ENTITY (ORIGINAL EVENT CODE: M1551); ENTITY STATUS OF PATENT OWNER: LARGE ENTITY

Year of fee payment: 4